 Patent Application Title |
Patent App Num. |
Date |
| Thermostable nucleic acid polymerase from thermococcus gorgonarius | 20110020898 | 20110127 |
| A purified thermostable enzyme is derived form the thermophilic archaebacterium Thermococcus gorgonarius. The enzyme can be native or recombinant, retains approximately 90% of its activity after incubation for two hours at 95° C. in the presence of stabilizing agents and possesses 3′-5′ proofreading exonuclease activity. Thermostable DNA polymerases are useful in many recombinant DNA techniques, especially nucleic acid amplification by the polymerase chain reaction (PCR).
... |
| Rna-dependent dna polymerase from geobacillus stearothermophilus | 20110020897 | 20110127 |
| The invention provides an isolated polynucleotide sequence from the genome of Bacillus stearothermophilus (Geobacillus stearothermophilus) and an amino acid sequence encoded by the polynucleotide sequence, the corresponding amino acid sequence comprising a novel enzyme, Tirt (thermostable intron reverse transcriptase), having reverse transcriptase activity and retaining that activity at temperatures of up to about 75° C.
... |
| Mutant dna polymerases and their genes | 20110020896 | 20110127 |
| The present invention relates to mutant DNA polymerases, their genes and their uses. More specifically, the present invention relates to mutant DNA polymerases which are originally isolated from Thermococcus sp NA1. strain and produced by site-specific mutagenesis, their amino acid sequences, genes encoding said mutant DNA polymerases, their nucleic acids and PCR methods by using thereof. As mutant DNA polymerases according to the present invention have decreased proofreading activity and changed function of inosine sensing effectively compared to wild type DNA polymerase, PCR using primers with specific nucleic acids has made rapid progress. Therefore, the present invention is broadly applicable for PCR in various molecular genetic technologies.
... |
| Method of removing nucleic acid contamination in reverse transcription and amplification reactions | 20110020878 | 20110127 |
| The invention provides methods of removing nucleic acid contamination from reverse transcription reactions and hot-start PCR, wherein said hot-start PCR is a barrier hot-start PCR set up and/or involves a hot-start DNA polymerase, which methods comprise use of a DNase that is substantially irreversibly inactivated by heating at a temperature of about 50° C. for 5 minutes, and that is substantially specific for double stranded DNA. The invention further provides a DNase that is substantially irreversibly inactivated by heating at a temperature of about 50° C. for 5 minutes, and that is substantially specific for double stranded DNA, nucleic acids encoding said DNase and kits or compositions comprising said DNase or said nucleic acid.
... |
| Rapid dna sequencing by peroxidative reaction | 20110020806 | 20110127 |
| Disclosed is a method of polynucleic acid (e.g., DNA) sequencing which is based on the generation of pyrophosphate (PPi) that occurs when a complementary base is incorporated into a growing DNA strand being synthesized on a template. The method utilizes a cascade of enzymatic reactions catalyzed by hypoxanthine-phosphoribosyl transferase, xanthine oxidase, and peroxidase in addition to DNA polymerase and apyrase. The last chemical step in the cascade of reactions is the oxidation of a material such as an electrode or luminol by hydrogen peroxide. This generates a detectable electrical or optical signal. This method is independent of luciferase, does not require dATP analogue, and is intended to improve precision and sensitivity of DNA sequencing, and to lessen the unsynchronized polymerization.
... |
| |
| Dna replication factors | 20110014679 | 20110120 |
| A DNA polymerase reaction system which provides high DNA polymerase activity even at a high temperature and at a high salt concentration. A DNA polymerase reaction system that is constructed from a DNA polymerase, a clamp, and a clamp loader without intein sequence, the DNA polymerase being from Pyrococcus horikoshii, a hyperthermophilic archaeon.
... |
| Thermostable dna polymerase from palaeococcus ferrophilus | 20110014660 | 20110120 |
| There is provided a polypeptide having thermostable DNA polymerase activity and comprising or consisting of an amino acid sequence with at least 90% identity to Palaeococcus ferrophilus DNA polymerase shown in SEQ ID NO: 1.
... |
| Method for preparing dna fragment having sticky end | 20110009607 | 20110113 |
| The present invention provides a method for preparing a DNA fragment, in which a desired double-stranded DNA fragment having a sticky end is directly and easily obtained from an amplification product (an amplified fragment) after PCR without a restriction enzyme digestion. The method for preparing a DNA fragment having a sticky end of the present invention comprises: (i) a step of performing a PCR reaction using a template DNA and specific primers to obtain an amplified DNA fragment; and (ii) a step of performing a prescribed treatment on the amplified DNA fragment to dissociate a protecting group from the fragment. Herein, the above-mentioned specific primers are composed of a complementary DNA portion consisting of a nucleotide sequence complementarily binding to an amplification target region in a... |
| Enzyme | 20110008848 | 20110113 |
| There is provided a polypeptide having thermostable DNA polymerase activity and comprising or consisting of an amino acid sequence with at least 55% identity to Thermodesulfatator indicus DNA polymerase I Large fragment shown in SEQ ID NO: 1 or in SEQ ID NO:32.
... |
| Mutant dna polymerases and methods of use | 20100323406 | 20101223 |
| The present invention provides mutant DNA polymerases, polynucleotides encoding the polymerases, cassettes and vectors including such polynucleotides, and cells containing the polymerases, polynucleotides, cassettes, and/or vectors of the invention. The present invention also provides methods for synthesizing polynucleotides and kits including a DNA polymerase of the invention.
... |
| Conditionally replicating viruses for cancer therapy | 20100316609 | 20101216 |
| Described herein are viral vectors comprising a nucleic acid encoding a viral DNA polymerase wherein the encoded viral DNA polymerase comprises at least one amino acid modification. Also provided are methods of making and using the viral vectors.
... |
| Dna polymerases having improved labeled nucleotide incorporation properties | 20100311959 | 20101209 |
| The present invention relates to mutant DNA polymerases that exhibit reduced discrimination against labeled nucleotides into polynucleotides. The DNA polymerases of the invention have at least one mutation in the nucleotide label interaction region of the enzyme such the mutation results in reduced discrimination against labeled nucleotides. The nucleotide label interaction regions is located at portions of the O-helix, (ii) the K helix, and (iii) the inter O—P helical loop of Taq DNA polymerase or analogous positions in other DNA polymerases. In addition to providing novel mutant DNA polymerases, the invention also provides polynucleotides encoding the subject mutant DNA polymerases. The polynucleotides provided may comprise expression vectors for the recombinant production of the mutant polymerases. The invention also provides host cells containing the subject polynucleotides. The... |
| Mutant dna polymerases | 20100311144 | 20101209 |
| Provided herein are mutant DNA-dependent polymerases which are derived from, or otherwise related to, wild type RB69 DNA polymerase. These mutant polymerases are capable of selectively binding labeled nucleotides. These mutant polymerases are also capable of incorporating a variety of naturally occurring and modified nucleotides, including, for example, terminator nucleotides.
... |
| |
| Recombinase polymerase amplification | 20100311127 | 20101209 |
| This disclosure describes related novel methods for Recombinase-Polymerase Amplification (RPA) of a target DNA that exploit the properties of recombinase and related proteins, to invade double-stranded DNA with single stranded homologous DNA permitting sequence specific priming of DNA polymerase reactions. The disclosed methods have the advantage of not requiring thermocycling or thermophilic enzymes, thus offering easy and affordable implementation and portability relative to other amplification methods. Further RPA reactions using light and otherwise, methods to determine the nature of amplified species without a need for gel electrophoresis, methods to improve and optimize signal to noise ratios in RPA reactions, methods to optimize oligonucleotide primer function, methods to control carry-over contamination, and methods to employ sequence-specific third ‘specificity’ probes. Further described are novel properties and approaches for... |
| Method for in vitro recombination | 20100311126 | 20101209 |
| The present invention relates, e.g., to an in vitro method, using isolated protein reagents, for joining two double-stranded (ds) DNA molecules of interest, wherein the distal region of the first DNA molecule and the proximal region of the second DNA molecule share a region of sequence identity, comprising (a) chewing back the DNA molecules with an enzyme having an exonuclease activity, to yield single-stranded overhanging portions of each DNA molecule which contain a sufficient length of the region of sequence identity to hybridize specifically to each other; (b) specifically annealing the single-stranded overhangs; and (c) repairing single-stranded gaps in the annealed DNA molecules and sealing the nicks thus formed (ligating the nicked DNA molecules). The region of sequence identity generally comprises at least 20 non-palindromic nucleotides... |
| Random-primed transcriptase -in vitro transcription method for rna amplification | 20100297728 | 20101125 |
| A random-primed reverse transcriptase-in vitro transcription method of linearly amplifying RNA is provided. According to the methods of the invention, source RNA (or other single-stranded nucleic acid), preferably, mRNA, is converted to double-stranded cDNA using two random primers, one of which comprises a RNA polymerase promoter sequence (“promoter-primer”), to yield a double-stranded cDNA that comprises a RNA polymerase promoter that is recognized by a RNA polymerase. Preferably, the primer for first-strand cDNA synthesis is a promoter-primer and the primer for second-strand cDNA synthesis is not a promoter-primer. The double-stranded cDNA is then transcribed into RNA by the RNA polymerase, optimally in the presence of a reverse transcriptase that is rendered incapable of RNA-dependent DNA polymerase activity during this transcription step. The subject methods produce linearly amplified... |
| Mutant dna polymerases and their genes from thermococcus | 20100297706 | 20101125 |
| The present invention relates to mutant DNA polymerases and their genes isolated from Thermococcus sp. More specifically, the present invention relates to mutant DNA polymerases which are originally isolated from Thermococcus sp NA1. strain and produced by site-specific mutagenesis, their amino acid sequences, genes encoding said mutant DNA polymerases, their nucleic acids sequences, recombinant vectors containing said nucleic acids sequences, host cells transformed with thereof and methods for producing mutant DNA polymerase protein by using thereof. As mutant DNA polymerases according to the present invention have increased processivity by site-specific mutagenesis on exonuclease active site compared to wild type DNA polymerase, the present invention is broadly applicable for PCR in various molecular genetic technologies.
... |
| Terminus-specific dna modification using random-sequence template oligonucleotides | 20100297643 | 20101125 |
| A method is provided for analyzing DNA molecules having unknown 3′ terminal sequences. The method involves contacting a DNA molecule with a plurality of template oligonucleotides blocked at their 3′ termini such that the template oligonucleotides are not extendable by DNA polymerase. The 3′ proximal portions of each of the template oligonucleotides comprise a region of random sequence and the 5′ proximal portions of each of the template oligonucleotides comprise the complement of a tag sequence. The DNA molecule and the template oligonucleotides are combined under conditions wherein the 3′ terminus of the DNA molecule hybridizes to the 3′ proximal portion of a template oligonucleotide and is extended by a DNA polymerase to produce a DNA molecule comprising a 3′ terminal tag sequence, and wherein the... |
| Thermostable dna polymerases and methods of use | 20100291638 | 20101118 |
| Thermostable viral and microbial polymerases exhibiting a combination of activities selected from proofreading (3′-5′) exonuclease activity, nick translating (5′-3′) nuclease activity, synthetic primer-initiated polymerase activity, nick-initiated polymerase activity, reverse transcriptase activity, strand displacement activity, terminal transferase activity, primase activity, and/or efficient incorporation of chain terminating analogs. Some of the polymerases provided herein include a first motif and a second motif. The first motif preferably has the sequence X1X2X3DX4PX5IELRX6X7X8, wherein X1 is I or V; X4 is F or Y; X8 is G or A; and X2, X3, X5, X6, and X7 are any amino acid. The second motif preferably has the sequence RX9X10X11KSANX12GX13X14YG, wherein X11 is G or A; X12 is F, L, or Y; X13 is L or V; X14 is I or L; and... |
| Combinatorial production of nucleotide and nucleoside (xitp) analogues | 20100286386 | 20101111 |
| Nucleotide analogues comprising a reactive hydrazide function (formula II) used as initial synthons for preparing compounds (formula I) capable of inducing mutations or capable of inhibiting a DNA polymerase or a kinase provided. Nucleic acids comprising the nucleotide and nucleoside analogues are also provided. Methods of using the compounds are also provided.
... |
| Method and apparatus for amplification of nucleic acid sequences by using thermal convection | 20100285536 | 20101111 |
| The present invention provides a nucleic acid sequence amplification method and apparatuses thereof that are simple in the design and easy to miniaturize and integrate into complex apparatuses, with capability of using DNA polymerases that are not thermostable. In the present invention, a plurality of heat sources are combined to supply or remove heat from specific regions of the sample such that a specific spatial temperature distribution is maintained inside the sample by locating a relatively high temperature region lower in height than a relatively low temperature region.
... |
| Markers associated with resistance to aphis glycines and methods of use therefor | 20100263085 | 20101014 |
| Methods for conveying Aphis glycines resistance (RAG) into non-resistant soybean germplasm are provided. In some embodiments, the methods include introgressing RAG into a non-resistant soybean using one or more nucleic acid markers for marker-assisted breeding among soybean lines to be used in a soybean breeding program, wherein the markers are linked to and/or associated with RAG. Also provided are single nucleotide polymorphisms (SNPs) associated with resistance to Aphis glycines; soybean plants, seeds, and tissue cultures produced by any of the disclosed methods; seed produced by the disclosed soybean plants; and compositions including amplification primer pairs capable of initiating DNA polymerization by a DNA polymerase on soybean nucleic acid templates to generate soybean marker amplicons.
... |
| Nucleic acid amplification method | 20100255546 | 20101007 |
| Disclosed is a novel isothermal nucleic acid amplification method enabling inexpensive and simple and easy detection. The method includes introducing nicking enzyme recognition sequences into an analysis target nucleic acid using nicking enzyme recognition sequence-containing primers and isothermally amplifying a specific region of the target nucleic acid using the primers, a nicking enzyme and a DNA polymerase having strand displacement activity.
... |
| Composition for analyzing nucleic acid | 20100255480 | 20101007 |
| The present invention relates to a composition for analyzing nucleic acid that contains luciferase for which reactivity to dATP is equal to or less than 1/400 reactivity to ATP, a method of analyzing nucleic acid that comprises the use of that composition, and a kit for analyzing nucleic acid comprising that composition.
... |
| F.oxysporum f.sp. melonis race 1,2-resistant melons | 20100235941 | 20100916 |
| Methods for conveying Fusarium oxysporum f.sp. melonis (FOM) race 1,2 resistance into non-resistant melon germplasm are provided, in some embodiments, the methods include introgressing FOM race 1,2 resistance into a non-resistant melon using one or more nucleic acid markers for marker-assisted selection among melon lines to be used in a melon breeding program, wherein the markers are linked to FOM race 1,2 resistance. Also provided are quantitative trait loci (QTLs) associated with resistance to FOM race 1,2; isolated and purified genetic markers associated with FOM race 1,2 resistance; melon plants, seeds, and tissue cultures produced by any of the disclosed methods; fruit and seed produced by the disclosed melon plants; and compositions including amplification primer pairs capable of initiating DNA polymerization by a DNA polymerase on... |
| Isothermal amplification method and dna polymerase used in the same | 20100221787 | 20100902 |
| (X) a primer that contains, in a 3′ end portion, a sequence (Ac′) that hybridizes to a sequence (A) of a 3′ end portion of the target nucleic acid sequence and also contains, on a 5′ side of the sequence (Ac′), a sequence (B′) that hybridizes to a complementary sequence (Bc) to a sequence (B) present on a 5′ side with respect to the sequence (A) in the target nucleic acid sequence
... |
| Cyclic reverse transcription method | 20100221786 | 20100902 |
| Disclosed is a cyclic reverse transcription method, which comprises performing reverse transcription reaction in one or more cycles, each cycle comprising the steps of: (i) performing reaction with a reverse transcription reaction solution comprising template RNA, RNA-dependent DNA polymerase, a reaction buffer, primers for reverse transcription and dNTPs, and optionally, a stabilizer, at 10° C. to 40° G and, (ii) performing reaction with the resultant reaction solution at 42° to 55° C.
... |
| Isothermal strand displacement amplification using primers containing a non-regular base | 20100221785 | 20100902 |
| The invention is directed to a method for isothermal DNA amplification comprising providing to the DNA to be amplified an amplification mix comprising a first primer at least partially complementary to a region of DNA and containing a non-regular base, a second primer at least partially complementary to a region of DNA and containing a non-regular base, a DNA polymerase, an enzyme capable of strand displacement, an enzyme that recognises a non-regular base in double-stranded DNA and causes a nick or excises a base in one DNA strand at or near the non-regular base; and amplifying the DNA substantially without thermal cycling.
... |
| Dried composition for hot-start pcr with long-term stability | 20100209973 | 20100819 |
| The present invention relates to a dried composition for hot-start PCR, more precisely a dried composition for hot-start PCR with improved stability and long-term storagability which is characteristically prepared by the steps of preparing a reaction mixture by mixing an aqueous solution containing reaction buffer, MgCl2, 4 types of dNTPs, DNA polymerase with pyrophosphate and pyrophosphatase in a reaction tube; and drying the reaction mixture prepared above, a preparation method of the same and a method for amplifying nucleic acid using the same. The dried composition for hot-start PCR is added with pyrophosphate and pyrophosphatase together before drying, so that it can have improved stability and long-term storagability as well as convenience in use, compared with the conventional compositions for hot-start PCR. Therefore, this composition can... |
| Method of amplification of gc-rich dna templates | 20100209970 | 20100819 |
| Methods are provided for increasing the processivity of DNA polymerases on GC-rich templates. The methods relate to providing enhancers and biased ratios of dNTPs, and may be used in DNA amplification reactions. The methods are useful for detecting genotypes associated with GC-rich repeats, including Fragile X Syndrome.
... |
| Dna replication factors | 20110014679 | 20110120 |
| A DNA polymerase reaction system which provides high DNA polymerase activity even at a high temperature and at a high salt concentration. A DNA polymerase reaction system that is constructed from a DNA polymerase, a clamp, and a clamp loader without intein sequence, the DNA polymerase being from Pyrococcus horikoshii, a hyperthermophilic archaeon.
... |
| Thermostable dna polymerase from palaeococcus ferrophilus | 20110014660 | 20110120 |
| There is provided a polypeptide having thermostable DNA polymerase activity and comprising or consisting of an amino acid sequence with at least 90% identity to Palaeococcus ferrophilus DNA polymerase shown in SEQ ID NO: 1.
... |
| Method for preparing dna fragment having sticky end | 20110009607 | 20110113 |
| The present invention provides a method for preparing a DNA fragment, in which a desired double-stranded DNA fragment having a sticky end is directly and easily obtained from an amplification product (an amplified fragment) after PCR without a restriction enzyme digestion. The method for preparing a DNA fragment having a sticky end of the present invention comprises: (i) a step of performing a PCR reaction using a template DNA and specific primers to obtain an amplified DNA fragment; and (ii) a step of performing a prescribed treatment on the amplified DNA fragment to dissociate a protecting group from the fragment. Herein, the above-mentioned specific primers are composed of a complementary DNA portion consisting of a nucleotide sequence complementarily binding to an amplification target region in a... |
| Enzyme | 20110008848 | 20110113 |
| There is provided a polypeptide having thermostable DNA polymerase activity and comprising or consisting of an amino acid sequence with at least 55% identity to Thermodesulfatator indicus DNA polymerase I Large fragment shown in SEQ ID NO: 1 or in SEQ ID NO:32.
... |
| Mutant dna polymerases and methods of use | 20100323406 | 20101223 |
| The present invention provides mutant DNA polymerases, polynucleotides encoding the polymerases, cassettes and vectors including such polynucleotides, and cells containing the polymerases, polynucleotides, cassettes, and/or vectors of the invention. The present invention also provides methods for synthesizing polynucleotides and kits including a DNA polymerase of the invention.
... |
| Conditionally replicating viruses for cancer therapy | 20100316609 | 20101216 |
| Described herein are viral vectors comprising a nucleic acid encoding a viral DNA polymerase wherein the encoded viral DNA polymerase comprises at least one amino acid modification. Also provided are methods of making and using the viral vectors.
... |
| Dna polymerases having improved labeled nucleotide incorporation properties | 20100311959 | 20101209 |
| The present invention relates to mutant DNA polymerases that exhibit reduced discrimination against labeled nucleotides into polynucleotides. The DNA polymerases of the invention have at least one mutation in the nucleotide label interaction region of the enzyme such the mutation results in reduced discrimination against labeled nucleotides. The nucleotide label interaction regions is located at portions of the O-helix, (ii) the K helix, and (iii) the inter O—P helical loop of Taq DNA polymerase or analogous positions in other DNA polymerases. In addition to providing novel mutant DNA polymerases, the invention also provides polynucleotides encoding the subject mutant DNA polymerases. The polynucleotides provided may comprise expression vectors for the recombinant production of the mutant polymerases. The invention also provides host cells containing the subject polynucleotides. The... |
| Mutant dna polymerases | 20100311144 | 20101209 |
| Provided herein are mutant DNA-dependent polymerases which are derived from, or otherwise related to, wild type RB69 DNA polymerase. These mutant polymerases are capable of selectively binding labeled nucleotides. These mutant polymerases are also capable of incorporating a variety of naturally occurring and modified nucleotides, including, for example, terminator nucleotides.
... |
| Recombinase polymerase amplification | 20100311127 | 20101209 |
| This disclosure describes related novel methods for Recombinase-Polymerase Amplification (RPA) of a target DNA that exploit the properties of recombinase and related proteins, to invade double-stranded DNA with single stranded homologous DNA permitting sequence specific priming of DNA polymerase reactions. The disclosed methods have the advantage of not requiring thermocycling or thermophilic enzymes, thus offering easy and affordable implementation and portability relative to other amplification methods. Further RPA reactions using light and otherwise, methods to determine the nature of amplified species without a need for gel electrophoresis, methods to improve and optimize signal to noise ratios in RPA reactions, methods to optimize oligonucleotide primer function, methods to control carry-over contamination, and methods to employ sequence-specific third ‘specificity’ probes. Further described are novel properties and approaches for... |
| Method for in vitro recombination | 20100311126 | 20101209 |
| The present invention relates, e.g., to an in vitro method, using isolated protein reagents, for joining two double-stranded (ds) DNA molecules of interest, wherein the distal region of the first DNA molecule and the proximal region of the second DNA molecule share a region of sequence identity, comprising (a) chewing back the DNA molecules with an enzyme having an exonuclease activity, to yield single-stranded overhanging portions of each DNA molecule which contain a sufficient length of the region of sequence identity to hybridize specifically to each other; (b) specifically annealing the single-stranded overhangs; and (c) repairing single-stranded gaps in the annealed DNA molecules and sealing the nicks thus formed (ligating the nicked DNA molecules). The region of sequence identity generally comprises at least 20 non-palindromic nucleotides... |
| Random-primed transcriptase -in vitro transcription method for rna amplification | 20100297728 | 20101125 |
| A random-primed reverse transcriptase-in vitro transcription method of linearly amplifying RNA is provided. According to the methods of the invention, source RNA (or other single-stranded nucleic acid), preferably, mRNA, is converted to double-stranded cDNA using two random primers, one of which comprises a RNA polymerase promoter sequence (“promoter-primer”), to yield a double-stranded cDNA that comprises a RNA polymerase promoter that is recognized by a RNA polymerase. Preferably, the primer for first-strand cDNA synthesis is a promoter-primer and the primer for second-strand cDNA synthesis is not a promoter-primer. The double-stranded cDNA is then transcribed into RNA by the RNA polymerase, optimally in the presence of a reverse transcriptase that is rendered incapable of RNA-dependent DNA polymerase activity during this transcription step. The subject methods produce linearly amplified... |
| Mutant dna polymerases and their genes from thermococcus | 20100297706 | 20101125 |
| The present invention relates to mutant DNA polymerases and their genes isolated from Thermococcus sp. More specifically, the present invention relates to mutant DNA polymerases which are originally isolated from Thermococcus sp NA1. strain and produced by site-specific mutagenesis, their amino acid sequences, genes encoding said mutant DNA polymerases, their nucleic acids sequences, recombinant vectors containing said nucleic acids sequences, host cells transformed with thereof and methods for producing mutant DNA polymerase protein by using thereof. As mutant DNA polymerases according to the present invention have increased processivity by site-specific mutagenesis on exonuclease active site compared to wild type DNA polymerase, the present invention is broadly applicable for PCR in various molecular genetic technologies.
... |
| Terminus-specific dna modification using random-sequence template oligonucleotides | 20100297643 | 20101125 |
| A method is provided for analyzing DNA molecules having unknown 3′ terminal sequences. The method involves contacting a DNA molecule with a plurality of template oligonucleotides blocked at their 3′ termini such that the template oligonucleotides are not extendable by DNA polymerase. The 3′ proximal portions of each of the template oligonucleotides comprise a region of random sequence and the 5′ proximal portions of each of the template oligonucleotides comprise the complement of a tag sequence. The DNA molecule and the template oligonucleotides are combined under conditions wherein the 3′ terminus of the DNA molecule hybridizes to the 3′ proximal portion of a template oligonucleotide and is extended by a DNA polymerase to produce a DNA molecule comprising a 3′ terminal tag sequence, and wherein the... |
| Thermostable dna polymerases and methods of use | 20100291638 | 20101118 |
| Thermostable viral and microbial polymerases exhibiting a combination of activities selected from proofreading (3′-5′) exonuclease activity, nick translating (5′-3′) nuclease activity, synthetic primer-initiated polymerase activity, nick-initiated polymerase activity, reverse transcriptase activity, strand displacement activity, terminal transferase activity, primase activity, and/or efficient incorporation of chain terminating analogs. Some of the polymerases provided herein include a first motif and a second motif. The first motif preferably has the sequence X1X2X3DX4PX5IELRX6X7X8, wherein X1 is I or V; X4 is F or Y; X8 is G or A; and X2, X3, X5, X6, and X7 are any amino acid. The second motif preferably has the sequence RX9X10X11KSANX12GX13X14YG, wherein X11 is G or A; X12 is F, L, or Y; X13 is L or V; X14 is I or L; and... |
| Combinatorial production of nucleotide and nucleoside (xitp) analogues | 20100286386 | 20101111 |
| Nucleotide analogues comprising a reactive hydrazide function (formula II) used as initial synthons for preparing compounds (formula I) capable of inducing mutations or capable of inhibiting a DNA polymerase or a kinase provided. Nucleic acids comprising the nucleotide and nucleoside analogues are also provided. Methods of using the compounds are also provided.
... |
| Method and apparatus for amplification of nucleic acid sequences by using thermal convection | 20100285536 | 20101111 |
| The present invention provides a nucleic acid sequence amplification method and apparatuses thereof that are simple in the design and easy to miniaturize and integrate into complex apparatuses, with capability of using DNA polymerases that are not thermostable. In the present invention, a plurality of heat sources are combined to supply or remove heat from specific regions of the sample such that a specific spatial temperature distribution is maintained inside the sample by locating a relatively high temperature region lower in height than a relatively low temperature region.
... |
| Markers associated with resistance to aphis glycines and methods of use therefor | 20100263085 | 20101014 |
| Methods for conveying Aphis glycines resistance (RAG) into non-resistant soybean germplasm are provided. In some embodiments, the methods include introgressing RAG into a non-resistant soybean using one or more nucleic acid markers for marker-assisted breeding among soybean lines to be used in a soybean breeding program, wherein the markers are linked to and/or associated with RAG. Also provided are single nucleotide polymorphisms (SNPs) associated with resistance to Aphis glycines; soybean plants, seeds, and tissue cultures produced by any of the disclosed methods; seed produced by the disclosed soybean plants; and compositions including amplification primer pairs capable of initiating DNA polymerization by a DNA polymerase on soybean nucleic acid templates to generate soybean marker amplicons.
... |
| Nucleic acid amplification method | 20100255546 | 20101007 |
| Disclosed is a novel isothermal nucleic acid amplification method enabling inexpensive and simple and easy detection. The method includes introducing nicking enzyme recognition sequences into an analysis target nucleic acid using nicking enzyme recognition sequence-containing primers and isothermally amplifying a specific region of the target nucleic acid using the primers, a nicking enzyme and a DNA polymerase having strand displacement activity.
... |
| Composition for analyzing nucleic acid | 20100255480 | 20101007 |
| The present invention relates to a composition for analyzing nucleic acid that contains luciferase for which reactivity to dATP is equal to or less than 1/400 reactivity to ATP, a method of analyzing nucleic acid that comprises the use of that composition, and a kit for analyzing nucleic acid comprising that composition.
... |
| F.oxysporum f.sp. melonis race 1,2-resistant melons | 20100235941 | 20100916 |
| Methods for conveying Fusarium oxysporum f.sp. melonis (FOM) race 1,2 resistance into non-resistant melon germplasm are provided, in some embodiments, the methods include introgressing FOM race 1,2 resistance into a non-resistant melon using one or more nucleic acid markers for marker-assisted selection among melon lines to be used in a melon breeding program, wherein the markers are linked to FOM race 1,2 resistance. Also provided are quantitative trait loci (QTLs) associated with resistance to FOM race 1,2; isolated and purified genetic markers associated with FOM race 1,2 resistance; melon plants, seeds, and tissue cultures produced by any of the disclosed methods; fruit and seed produced by the disclosed melon plants; and compositions including amplification primer pairs capable of initiating DNA polymerization by a DNA polymerase on... |
| Isothermal amplification method and dna polymerase used in the same | 20100221787 | 20100902 |
| (X) a primer that contains, in a 3′ end portion, a sequence (Ac′) that hybridizes to a sequence (A) of a 3′ end portion of the target nucleic acid sequence and also contains, on a 5′ side of the sequence (Ac′), a sequence (B′) that hybridizes to a complementary sequence (Bc) to a sequence (B) present on a 5′ side with respect to the sequence (A) in the target nucleic acid sequence
... |
| Cyclic reverse transcription method | 20100221786 | 20100902 |
| Disclosed is a cyclic reverse transcription method, which comprises performing reverse transcription reaction in one or more cycles, each cycle comprising the steps of: (i) performing reaction with a reverse transcription reaction solution comprising template RNA, RNA-dependent DNA polymerase, a reaction buffer, primers for reverse transcription and dNTPs, and optionally, a stabilizer, at 10° C. to 40° G and, (ii) performing reaction with the resultant reaction solution at 42° to 55° C.
... |
| Isothermal strand displacement amplification using primers containing a non-regular base | 20100221785 | 20100902 |
| The invention is directed to a method for isothermal DNA amplification comprising providing to the DNA to be amplified an amplification mix comprising a first primer at least partially complementary to a region of DNA and containing a non-regular base, a second primer at least partially complementary to a region of DNA and containing a non-regular base, a DNA polymerase, an enzyme capable of strand displacement, an enzyme that recognises a non-regular base in double-stranded DNA and causes a nick or excises a base in one DNA strand at or near the non-regular base; and amplifying the DNA substantially without thermal cycling.
... |
| Dried composition for hot-start pcr with long-term stability | 20100209973 | 20100819 |
| The present invention relates to a dried composition for hot-start PCR, more precisely a dried composition for hot-start PCR with improved stability and long-term storagability which is characteristically prepared by the steps of preparing a reaction mixture by mixing an aqueous solution containing reaction buffer, MgCl2, 4 types of dNTPs, DNA polymerase with pyrophosphate and pyrophosphatase in a reaction tube; and drying the reaction mixture prepared above, a preparation method of the same and a method for amplifying nucleic acid using the same. The dried composition for hot-start PCR is added with pyrophosphate and pyrophosphatase together before drying, so that it can have improved stability and long-term storagability as well as convenience in use, compared with the conventional compositions for hot-start PCR. Therefore, this composition can... |
| Method of amplification of gc-rich dna templates | 20100209970 | 20100819 |
| Methods are provided for increasing the processivity of DNA polymerases on GC-rich templates. The methods relate to providing enhancers and biased ratios of dNTPs, and may be used in DNA amplification reactions. The methods are useful for detecting genotypes associated with GC-rich repeats, including Fragile X Syndrome.
... |
| Method of determining the nucleotide sequence of oligonucleotides and dna molecules | 20100209922 | 20100819 |
| The present invention relates to a novel method for analyzing nucleic acid sequences based on real-time detection of DNA polymerase-catalyzed incorporation of each of the four nucleotide bases, supplied individually and serially in a microfluidic system, to a reaction cell containing a template system comprising a DNA fragment of unknown sequence and an oligonucleotide primer. Incorporation of a nucleotide base into the template system can be detected by any of a variety of methods including but not limited to fluorescence and chemiluminescence detection. Alternatively, microcalorimetic detection of the heat generated by the incorporation of a nucleotide into the extending template system using thermopile, thermistor and refractive index measurements can be used to detect extension reactions.
... |
| Recombinant reverse transcriptases | 20100203597 | 20100812 |
| The present invention relates to a gene that encodes a hyperactive reverse transcriptase having DNA polymerase activity and substantially reduced RNase H activity, vectors containing the gene and host cells transformed with the invention. The present invention also includes a method of producing the hyperactive reverse transcriptase, producing cDNA from mRNA using the reverse transcriptase of the invention, kits and assay templates made using the hyperactive reverse transcriptase.
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| High-speed pcr using high-speed dna polymerase | 20100203594 | 20100812 |
| [Means for Solving Problems] A method for practicing a high-speed PCR under a temperature change of 10° C./sec or more, characterized in that use is made of a heat-resistant DNA polymerase exhibiting a speed of synthesizing a deoxyribonucleic acid of 100 base/sec or higher, and a reagent kit for a high-speed PCR, or use in the method.
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| Ambient temperature stable kits for molecular diagnostics | 20100196904 | 20100805 |
| A method for processing DNA polymerase and/or dNTPs for use in an amplification procedure, includes providing a solution mixture, the solution mixture including a DNA polymerase and/or dNTPs, a buffer solution and at least one stabilizing agent and hydration reducing the solution mixture. The solution mixture is hydration reduced at a temperature between 0° C. and about 100° C.
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| Markers associated with soybean rust resistance and methods of use therefor | 20100192247 | 20100729 |
| Methods for conveying soybean rust (SBR) resistance into non-resistant soybean germplasm are provided. In some embodiments, the methods include introgressing SBR resistance into a non-resistant soybean using one or more nucleic acid markers for marker-assisted breeding among soybean lines to be used in a soybean breeding program, wherein the markers are linked to and/or associated with SBR resistance. Also provided are single nucleotide polymorphisms (SNPs) associated with resistance to SBR; soybean plants, seeds, and tissue cultures produced by any of the disclosed methods; seed produced by the disclosed soybean plants; and compositions including amplification primer pairs capable of initiating DNA polymerization by a DNA polymerase on soybean nucleic acid templates to generate soybean marker amplicons.
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| Thermoactive sivagm sab reverse transcriptase | 20090325235 | 20091231 |
| Methods and kits performing reverse transcription and RT-PCR reactions having high fidelity, processivity and DNA polymerase activity are described. The methods involve performance of reverse transcription at an increased temperature with a reverse transcriptase from Simian Immunodeficiency Virus-agm.sab or a variation thereof. The kits of the present invention include a reverse transcriptase from Simian Immunodeficiency Virus-agm.sab or a variation thereof, a DNA polymerase capable of amplifying cDNA under conditions suitable for polymerase chain reaction, and the reagents necessary to carry out both processes.
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| Recombinase polymerase amplification | 20090325165 | 20091231 |
| This disclosure describe three related novel methods for Recombinase-Polymerase Amplification (RPA) of a target DNA that exploit the properties of the bacterial RecA and related proteins, to invade double-stranded DNA with single stranded homologous DNA permitting sequence specific priming of DNA polymerase reactions. The disclosed methods has the advantage of not requiring thermocycling or thermophilic enzymes. Further, the improved processivity of the disclosed methods allow amplification of DNA up to hundreds of megabases in length.
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| Thermus egertssonii dna polymerases | 20090317888 | 20091224 |
| The present invention relates to a thermophilic polymerase, wherein the DNA polymerase has an in-vitro primer extension rate that is >35 bases/second and faster relative to the primer extension rate of a DNA polymerase comprising amino acid sequences SEQ ID NO: 2 or 4, when measured under identical conditions in a DNA replication assay using primed single strand M13mp18 DNA and an incubation temperature of 60° C. The invention also relates to a vector comprising the polymerase, a host cell comprising the vector. The invention relates to a nucleic acid replication kit comprising the polymerase according to the invention.
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| Method for amplifying genomic dna | 20090311753 | 20091217 |
| A method for amplifying genomic DNA is provided. The method comprises the steps of: (1) incubating a cell-containing agarose solution at a pH of 9 to 12 and a temperature of 45 to 80° C. to produce a genomic DNA-dispersed agarose solution wherein 0.002 to 1 copies/5 microliter of single-stranded genomic DNA is dispersed; (2) solidifying the genomic DNA-dispersed agarose solution to produce a genomic DNA-dispersed agarose gel and neutralizing a pH of the gel; and (3) adding a DNA polymerase with strand displacement activity, primer and dNTP to the genomic DNA-dispersed agarose gel and incubating the gel at a temperature of 0 to 60° C. to amplify the genomic DNA.
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| Polymerase | 20090305345 | 20091210 |
| The present invention relates to an engineered polymerase characterized in that the polymerase exhibits an enhanced ability to process nucleic acid in the presence of environmental and biological inhibitors compared to wild type DNA polymerase.
... |
| Dna polymerase | 20090305292 | 20091210 |
| The present invention relates to DNA polymerases. In particular the invention relates to a method for the generation of DNA polymerases exhibiting a relaxed substrate specificity. Uses of mutant polymerases produced using the methods of the invention are also described.
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| Method for synthesizing nucleic acid using dna polymerase beta and single molecule sequencing method | 20090291440 | 20091126 |
| The present invention provides a nucleic acid synthesis method capable of continuously carrying out an extension reaction and a single molecule sequencing method capable of obtaining base information accurately at high speed. A method for synthesizing a nucleic acid, including the steps of: forming a complex of a target nucleic acid hybridized to a primer oligonucleotide and a DNA polymerase β; allowing the DNA polymerase β to incorporate a fluorescently-labeled dNTP so that the fluorescently-labeled dNTP is bound to the 3′ end of the primer oligonucleotide; and allowing the DNA polymerase β to continuously incorporate fluorescently-labeled dNTP to extend a nucleic acid complementary to the target nucleic acid from the 3′ end of the bound fluorescently-labeled dNTP. A method for sequencing a single nucleic acid molecule,... |
| Novel nucleic acid base pair | 20090286247 | 20091119 |
| A novel artificial nucleic acid base pair which is obtained by forming a selective base pair by introducing a group having steric hindrance (preferably a group having steric hindrance and static repulsion and a stacking effect) and can be recognized by a polymerase such as DNA polymerase; a novel artificial gene; and a method of designing nucleic acid bases so as to form a selective base pair with the use of steric hindrance, static repulsion and stacking effect at the base moiety of the nucleic acid. An artificial nucleic acid comprising these bases; a process for producing the same; a codon containing the same; a nucleic acid molecule containing the same; a process for producing a non-natural gene by using the same; a process for producing... |
| Dna polymerases and related methods | 20090280539 | 20091112 |
| Disclosed are mutant DNA polymerases having improved extension rates relative to a corresponding, unmodified polymerase. The mutant polymerases are useful in a variety of disclosed primer extension methods. The mutant polymerases overcome the inhibitory effects by an intercalating dye. Therefore, the mutant polymerases are useful in a variety of disclosed methods in combination with an intercalating dye. Also disclosed are related compositions, including recombinant nucleic acids, vectors, and host cells, which are useful, e.g., for production of the mutant DNA polymerases.
... |
| Dna polymerases and related methods | 20090280539 | 20091112 |
| Disclosed are mutant DNA polymerases having improved extension rates relative to a corresponding, unmodified polymerase. The mutant polymerases are useful in a variety of disclosed primer extension methods. The mutant polymerases overcome the inhibitory effects by an intercalating dye. Therefore, the mutant polymerases are useful in a variety of disclosed methods in combination with an intercalating dye. Also disclosed are related compositions, including recombinant nucleic acids, vectors, and host cells, which are useful, e.g., for production of the mutant DNA polymerases.
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| Nucleic acid amplification in the presence of modified randomers | 20090269766 | 20091029 |
| The present invention is directed to a composition comprising a DNA Polymerase which is preferably thermostable, Deoxynucleotides, at least one primer oligonucleotide or a pair of amplification primers, and randomized 5-8 mer oligonucleotide, characterized in that said oligonucleotide comprises a modification with an organic hydrophobic moiety Such a composition is specifically useful for performing hot start PCR.
... |
| Nucleic acid amplification in the presence of modified randomers | 20090269766 | 20091029 |
| The present invention is directed to a composition comprising a DNA Polymerase which is preferably thermostable, Deoxynucleotides, at least one primer oligonucleotide or a pair of amplification primers, and randomized 5-8 mer oligonucleotide, characterized in that said oligonucleotide comprises a modification with an organic hydrophobic moiety Such a composition is specifically useful for performing hot start PCR.
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| Methods and compositions for pcr | 20090263869 | 20091022 |
| A modified thermostable Pol B DNA polymerase, produced by a reaction, under essentially aqueous conditions, of a thermostable Pol B DNA polymerase and a modifier reagent of Formula I wherein the reaction results in a thermally reversible inactivation of the thermostable Pol B DNA polymerase activity and the 3′-5′ exonuclease activity, which polymerase is suitable for hot-start PCR. Also disclosed are the method for the modification, a polynucleic acid amplification method and PCR reaction mixture and kit comprising the modified thermostable Pol B DNA polymerase.
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| Methods and compositions for amplification of dna | 20090263871 | 20091022 |
| The invention provides an Enzyme Blend comprising a DNA polymerase and a DNA repair enzyme. Methods and kits for amplification of DNA that is damaged, undamaged, or suspected of being damaged are also provided.
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| Methods of rna amplification in the presence of dna | 20090239232 | 20090924 |
| The invention provides methods for amplification of RNA. The methods are particularly suitable for specifically amplifying RNA in the presence of DNA. The methods involve producing a marked first primer extension product from a target RNA in the presence of a DNA-dependent DNA polymerase inhibitor, which prevents replication of DNA by the reverse transcriptase enzyme. The marked nucleic acid products are subsequently selectively amplified in the presence on non-marked nucleic acids. The methods are useful for production and analysis of polynucleotide sequences complementary to an RNA sequence. The methods are useful for preparation of nucleic acid libraries and substrates for analysis of gene expression of cells in biological samples. The invention also provides compositions and kits for practicing the amplification methods, as well as methods which... |
| Pyrophosphorolysis activated polymerization (pap) | 20090239283 | 20090924 |
| A novel method of pyrophosphorolysis activated polymerization (PAP) has been developed. In PAP, pyrophosphorolysis and polymerization by DNA polymerase are coupled serially for each amplification by using an activatable oligonucleotide P* that has a non-extendible 3′-deoxynucleotide at its 3′ terminus. PAP can be applied for exponential amplification or for linear amplification. PAP can be applied to amplification of a rare allele in admixture with one or more wild-type alleles by using an activatable oligonucleotide P* that is an exact match at its 3′ end for the rare allele but has a mismatch at or near its 3′ terminus for the wild-type allele. PAP is inhibited by a mismatch in the 3′ specific sequence as far as 16 nucleotides away from the 3′ terminus. PAP can greatly... |
| Stable compositions for nucleic acid amplification and sequencing | 20090233283 | 20090917 |
| The present invention is directed to compositions comprising mixtures of reagents, including thermostable enzymes (e.g., thermostable DNA polymerases), buffers, cofactors and other components, suitable for immediate use in nucleic acid amplification or sequencing techniques without dilution or addition of further components. The compositions contain no stabilizing agents (e.g., glycerol or serum albumin) and unexpectedly maintain activity for extended periods of time upon storage at temperatures above freezing. These compositions are useful, alone or in the form of kits, for nucleic acid amplification (e.g., by the Polymerase Chain Reaction) and sequencing (e.g., by dideoxy or “Sanger” sequencing), or for any procedure utilizing thermostable DNA polymerases in a variety of medical, forensic and agricultural applications. In particular, the compositions and methods are useful for amplifying and sequencing nucleic... |
| Stable compositions for nucleic acid amplification and sequencing | 20090233283 | 20090917 |
| The present invention is directed to compositions comprising mixtures of reagents, including thermostable enzymes (e.g., thermostable DNA polymerases), buffers, cofactors and other components, suitable for immediate use in nucleic acid amplification or sequencing techniques without dilution or addition of further components. The compositions contain no stabilizing agents (e.g., glycerol or serum albumin) and unexpectedly maintain activity for extended periods of time upon storage at temperatures above freezing. These compositions are useful, alone or in the form of kits, for nucleic acid amplification (e.g., by the Polymerase Chain Reaction) and sequencing (e.g., by dideoxy or “Sanger” sequencing), or for any procedure utilizing thermostable DNA polymerases in a variety of medical, forensic and agricultural applications. In particular, the compositions and methods are useful for amplifying and sequencing nucleic... |
| Nucleic acid-free thermostable enzymes and methods of production thereof | 20090215126 | 20090827 |
| The present invention provides thermostable enzymes, such as DNA polymerases and restriction endonucleases, that are substantially free from contamination with nucleic acids. The invention also provides methods for the production of these enzymes, and kits comprising these enzymes which may be used in amplifying or sequencing nucleic acid molecules, including through use of the polymerase chain reaction (PCR).
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| Nucleic acid-free thermostable enzymes and methods of production thereof | 20090215126 | 20090827 |
| The present invention provides thermostable enzymes, such as DNA polymerases and restriction endonucleases, that are substantially free from contamination with nucleic acids. The invention also provides methods for the production of these enzymes, and kits comprising these enzymes which may be used in amplifying or sequencing nucleic acid molecules, including through use of the polymerase chain reaction (PCR).
... |
| Nucleic acid-free thermostable enzymes and methods of production thereof | 20090215126 | 20090827 |
| The present invention provides thermostable enzymes, such as DNA polymerases and restriction endonucleases, that are substantially free from contamination with nucleic acids. The invention also provides methods for the production of these enzymes, and kits comprising these enzymes which may be used in amplifying or sequencing nucleic acid molecules, including through use of the polymerase chain reaction (PCR).
... |
| Nucleic acid-free thermostable enzymes and methods of production thereof | 20090215126 | 20090827 |
| The present invention provides thermostable enzymes, such as DNA polymerases and restriction endonucleases, that are substantially free from contamination with nucleic acids. The invention also provides methods for the production of these enzymes, and kits comprising these enzymes which may be used in amplifying or sequencing nucleic acid molecules, including through use of the polymerase chain reaction (PCR).
... |
| Nucleic acid-free thermostable enzymes and methods of production thereof | 20090215126 | 20090827 |
| The present invention provides thermostable enzymes, such as DNA polymerases and restriction endonucleases, that are substantially free from contamination with nucleic acids. The invention also provides methods for the production of these enzymes, and kits comprising these enzymes which may be used in amplifying or sequencing nucleic acid molecules, including through use of the polymerase chain reaction (PCR).
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| Chimeric dna polymerase | 20090209005 | 20090820 |
| The present invention provides a chimeric thermostable DNA polymerase that includes a region from a Tth DNA polymerase I, a region from a Taq DNA polymerase I and a DNA polymerase domain. The DNA polymerase domain comprises a portion of a DNA polymerase domain from the Tth DNA polymerase I operably linked to a portion of a DNA polymerase domain from the Taq DNA polymerase I. Also provided are a nucleic acid sequence and an amino acid sequence of the chimeric thermostable enzyme of the invention. The chimeric DNA polymerase enzyme of the invention is useful in DNA amplification reactions such as the polymerase chain reaction.
... |
| Chimeric dna polymerase | 20090209005 | 20090820 |
| The present invention provides a chimeric thermostable DNA polymerase that includes a region from a Tth DNA polymerase I, a region from a Taq DNA polymerase I and a DNA polymerase domain. The DNA polymerase domain comprises a portion of a DNA polymerase domain from the Tth DNA polymerase I operably linked to a portion of a DNA polymerase domain from the Taq DNA polymerase I. Also provided are a nucleic acid sequence and an amino acid sequence of the chimeric thermostable enzyme of the invention. The chimeric DNA polymerase enzyme of the invention is useful in DNA amplification reactions such as the polymerase chain reaction.
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| Use of dna polymerases | 20090202993 | 20090813 |
| Use of a DNA polymerase enzyme as a single stranded RNA exoribonuclease.
... |
| Mutant dna polymerases and uses therof | 20090191560 | 20090730 |
| The present invention relates to mutant DNA polymerases which incorporate dideoxynucleotides with about the same efficiency as deoxynucleotides. The present invention also related to mutant DNA polymerases which also have substantially reduced 5′-to-3′ exonuclease activity or 3′-to-5′ exonuclease activity. The invention also relates to DNA molecules coding for the mutant DNA polymerases, and hosts containing the DNA molecules.
... |
| Method for amplification of long nucleic acid | 20090191595 | 20090730 |
| An object of the present invention is to provide a method for amplification of long nucleic acid, wherein the method allows nucleic acid fragments containing the same nucleotide sequence information to efficiently amplify at the same base length. The present invention relates to a method for amplification of long nucleic acid sequence, wherein the method uses primers being modified at the 5′ end with a phosphate group and performs a cooperative reaction using DNA polymerase and DNA ligase.
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| Detection format for hot start real time polymerase chain reaction | 20090181401 | 20090716 |
| The present invention is directed to a method and a composition for amplifying and detecting a target nucleic comprising subjecting said target nucleic acid to a real time PCR amplification reaction in the presence of a thermostable DNA polymerase, a thermostable double strand dependent 3′-5′ exonuclease having a temperature optimum above 37° C., a pair of amplification primers, deoxynucleoside triphosphates, a detecting oligonucleotide carrying a first label and a second label, said first label being capable of acting as a fluorescent reporter entity when excited with light of an appropriate wavelength, said second label being capable of acting as a fluorescence quenching entity of said fluorescent reporter entity, characterized in that one label is bound to the 3′ end of said detecting oligonucleotide, and further characterized... |
| Amplification and cloning of single dna molecules using rolling circle amplification | 20090176280 | 20090709 |
| The present invention relates, e.g., to a method for amplifying a small number of copies (e.g. a single copy) of a single-stranded circular DNA molecule (e.g. having a size of about 5-6 kb) by an isothermal rolling circle mechanism, using random or partially random primers and a F29-type DNA polymerase. The method, which can also be used for amplifying DNAs by non-rolling types of multiple displacement amplification, comprises incubating the reaction components in a small volume, e.g. about 10 μl or less, such as about 0.6 μl or less. The degree of amplification can be about 109 fold, or higher. A method for cell-free cloning of DNA, using the rolling circle amplification method of the invention, is described.
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| Apparatus, methods and products for detecting genetic mutation | 20080318215 | 20081225 |
| Methods for detecting genetic mutation allowing detection of very low frequency mutation. Methods comprise treating RNA:DNA heteroduplexes of interest with ribonuclease treatment coupled with DNA polymerase treatment. RNA:DNA heteroduplexes of interest are preferentially targeted for digestion by ribonuclease and subsequent sequence extension by DNA polymerase. Methods may be carried out partially or entirely manually, automatically, and combinations thereof. Methods may be performed wholly or partially in solution, on solid phase media, in large scale, adapted for high throughput analysis, and any combinations thereof. Apparatus and products for detecting genetic mutation.
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| Thermostabillization of dna polymerase by protein folding pathway from a hyperthermophile archaeon, pyrococcus furiosus | 20080318281 | 20081225 |
| The present invention relates to maintaining the activity and stability of enzymes and biologically active proteins at increased temperatures by contacting same with a combination of isolated passive and active chaperones from a hyperthermopilic Archaeon, wherein the chaperones may include heat shock proteins, prefoldin and/or chaperonin proteins.
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| Dna polymerases having strand displacement activity | 20080311626 | 20081218 |
| The invention provides novel strand displacement DNA polymerases which can be used in a rapid and efficient strand displacement amplification reactions. The polymerases are significantly more thermostable than prior art polymerase and retain high activity at elevated temperatures. Also disclosed are genes encoding the polymerases and vectors comprising the genes. Representative polymerases of the invention are obtainable from bacterial strains of the species Thermus antranikianii and Thermus brockianus and also from environmental samples with isolation of the source species.
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| Compositions and methods for preventing carry-over contamination in nucleic acid amplification reactions | 20080311627 | 20081218 |
| Particular aspects provide methods for the specific amplification of template DNA in the presence of potentially contaminating PCR products from previous amplification experiments, but wherein the contaminating prior reaction amplificates (carry-over contaminants) are rendered non-amplifiable, based on use of particular contaminant degradation enzymes, and use of at least one primer that is fully complementary to any contaminating prior reaction amplificate nucleic acid, but contains a mismatch with the correspond sequence of the sample template nucleic acid. Additional aspects provide a method for the specific amplification of single-stranded sample template DNA in the presence of potentially double-stranded carry-over products. Further aspects provide methods comprising use of a template-dependent thermostable DNA polymerase enzyme suitable for incorporating ribonucleotides as well as deoxy-nucleotides to provide a chimeric amplificate. After digestion... |
| Recombinase polymerase amplification | 20080293045 | 20081127 |
| This disclosure describes related novel methods for Recombinase-Polymerase Amplification (RPA) of a target DNA that exploit the properties of recombinase and related proteins, to invade double-stranded DNA with single stranded homologous DNA permitting sequence specific priming of DNA polymerase reactions. The disclosed methods have the advantage of not requiring thermocycling or thermophilic enzymes, thus offering easy and affordable implementation and portability relative to other amplification methods. Further disclosed are conditions to enable real-time monitoring of RPA reactions, methods to regulate RPA reactions using light and otherwise, methods to determine the nature of amplified species without a need for gel electrophoresis, methods to improve and optimize signal to noise ratios in RPA reactions, methods to optimize oligonucleotide primer function, methods to control carryover contamination, and methods to... |
| Method of analyzing dna sequence using field-effect device, and base sequence analyzer | 20080286767 | 20081120 |
| Since the elongation reaction of one base induced at the gate portion can be directly converted to an electrical signal, expensive lasers or complex optical systems are not needed. Thus, a small gene polymorphism detection system that can conduct measurement at high precision can be provided.
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| Compositions and methods utilizing dna polymerases | 20080280291 | 20081113 |
| The invention features a novel isolated Family B DNA polymerase, a Thermococcus polymerase JDF-3, and mutant recombinant forms thereof. Mutant polymerases of the invention are deficient in 3′ to 5′ exonuclease activity and/or exhibit reduced discrimination against non-conventional nucleotides relative to the wild-type form of the polymerase.
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| Antiviral resistance mutants and applications thereof | 20080274083 | 20081106 |
| The present invention relates generally to viral variants exhibiting reduced sensitivity to agents such as nucleoside or nucleotide analogs or other DNA polymerase antagonists and/or reduced interactivity with immunological reagents. More particularly, the present invention is directed to hepatitis B virus (HBV) variants exhibiting complete or partial resistance to nucleoside or nucleotide analogs or other DNA polymerase antagonists and/or reduced interactivity with antibodies to viral surface components including reduced sensitivity to these antibodies. The present invention further contemplates assays for detecting such viral variants, which assays are useful in monitoring anti-viral therapeutic regimens and in developing new or modified vaccines directed against viral agents and in particular HBV variants. The present invention also contemplates the use of the viral variants to screen for and/or develop or... |
| Methods and kits for negative selection of desired nucleic acid sequences | 20080268508 | 20081030 |
| The present invention pertains to a method to isolate, separate, enrich or amplify a targeted nucleotide polymer such as mRNA through selective reverse transcription of the targeted polymer into cDNA from a sample comprising of chemically identical or similar polynucleotide polymers such as rRNA. The enrichment of the targeted nucleic acid such as mRNA is accomplished by blocking the reverse transcription of undesired rRNA while allowing unrestricted reverse transcription of the targeted polymer. The invention also embodies that the cleavage of the non-targeted nucleic acid such as rRNA bound to an oligonucleotide through enzymatic activity (RNase H). The invention further embodies methods and kits to accomplish the utility of the invention through the following steps 1) 3′ tailing of chemically identical or similar nucleotide polymers in... |
| Method and a system for predicting protein functional site, a method for improving protein function, and a function-modified protein | 20080262201 | 20081023 |
| The present application provides a method for predicting the functional site of a protein using data of the entire proteins of an organism of which genome data or cDNA data is known. More specifically, the present application provides a method for predicting a protein functional site, comprising the steps of calculating the frequency of occurrence of an oligopeptide in the entire proteins, calculating the value of each amino-acid residue contributing to the frequency of occurrence as the representative value of the function, and predicting the protein functional site by using the representative value of function as an indicator. The present also provides a system for predicting a functional site for automatically performing said methods. Additionally, the present application provides a method for preparing a function-modified protein... |
| Dna polymerase blends and mutant dna polymerases | 20080254525 | 20081016 |
| A thermostable DNA polymerase composition comprising at least two DNA polymerases, one of which is substantially reduced in 5′-exonuclease activity and one of which has 5′-exonuclease activity. This polymerase may be used in methods including, but not limited to, nucleic acid synthesis, DNA sequencing, nucleic acid amplification and cDNA synthesis,
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| Method for the amplification and detection of hbv dna using a transcription based amplification | 20080241891 | 20081002 |
| The present invention provides a method for the transcription based amplification of a target HBV nucleic acid sequence starting from HBV DNA optionally present in a sample, comprising the steps of, —incubating the sample, suspected to contain HBV, in an amplification buffer with one or more restriction enzymes capable of cleaving the HBV DNA at a selected restriction site, said restriction enzyme creating a defined 3′ end of said HBV DNA stand(s), a promoter-primer, said promoter-primer having a 5′ region comprising the sequence of a promoter recognized by a DNA-dependent RNA polymerase and a 3′ region complementary to the define 3′ end of the DNA strand, a second or reverse primer, having the opposite polarity of the promoter-primer and comprising the 5′ end of the said... |
| Pna probes, kits, and methods for detecting lamivudine-resistant hepatitis b viruses | 20080233557 | 20080925 |
| Disclosed are peptide nucleic acid (PNA) probes to detect lamivudine resistant mutants of hepatitis B virus (HBV), which causes acute and chronic hepatitis, kits for detecting lamivudine resistant HBV comprising the PNA probes, and methods for detecting lamivudine resistant HBV by using the kits. They can accurately detect mutations of rtL180M, rtM204V, rtM204I and rtV2071 within B and C domains of HBV DNA polymerase gene, the main cause of lamivudine resistance, as well as mixed mutants of more than one mutant. They can detect lamivudine resistant HBV with high specificity and sensitivity.
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| Detection method of dna amplification using probe labeled with intercalating dye | 20080220415 | 20080911 |
| The present invention relates to a detection method of nucleic acid amplification using probe labeled with intercalating dye. More particularly, the present invention is directed to a real-time detection method of nucleic acid amplification, comprising the steps of i) producing an aqueous buffer which contains a nucleic acid, a pair of primers for amplification of said nucleic acid, a fluorescent probe wherein a fluorescent dye of which intensity of fluorescence is varied when the dye is intercalated into a double-stranded nucleic acid, is connected with an oligonucleotide of which base sequence is complementary with at least a part of said nucleic acid, four (4) kinds of nucleotides and DNA polymerase; ii) denaturing said doublestranded nucleic acid into single strands by heating the aqueous buffer prepared in... |
| Method of determining the nucleotide sequence of oligonucleotides and dna molecules | 20080213770 | 20080904 |
| The present invention relates to a novel method for analyzing nucleic acid sequences based on real-time detection of DNA polymerase-catalyzed incorporation of each of the four nucleotide bases, supplied individually and serially in a microfluidic system, to a reaction cell containing a template system comprising a DNA fragment of unknown sequence and an oligonucleotide primer. Incorporation of a nucleotide base into the template system can be detected by any of a variety of methods including but not limited to fluorescence and chemiluminescence detection. Alternatively, microcalorimetic detection of the heat generated by the incorporation of a nucleotide into the extending template system using thermopile, thermistor and refractive index measurements can be used to detect extension reactions.
... |
| Methods for detecting and localizing dna mutations by microarray | 20080206875 | 20080828 |
| This disclosure provides methods for detecting and localizing DNA mutations by DNA microarray. In various embodiments, the described methods include use of restriction endonuclease(s) and/or mismatch-recognition nuclease(s) to detect and/or localize mutations. In one representative method, reference and target DNA are digested using one or more restriction endonucleases, resultant DNA strands are labeled (e.g., using a DNA polymerase), and the labeled mixture of DNAs is hybridized to a microarray. In another representative method, reference and target DNA are denatured and annealed to form a mixture containing heteroduplex DNA, one or more mismatch-recognition nuclease(s) are used to nick or cleave at least a portion of the heteroduplex DNA, resultant DNA strands are labeled (e.g., using a DNA polymerase) and the labeled mixture of DNAs is hybridized to... |
| Compositions and methods utilizing dna polymerases | 20080199935 | 20080821 |
| The invention features a novel isolated Family B DNA polymerase, a Thermococcus polymerase JDF-3, and mutant recombinant forms thereof. Mutant polymerases of the invention are deficient in 3′ to 5′ exonuclease activity and/or exhibit reduced discrimination against non-conventional nucleotides relative to the wild-type form of the polymerase.
... |
| Specialized oligonucleotides and their use in nucleic acid amplification and detection | 20080193934 | 20080814 |
| Described herein are labeled probes and unlabeled oligonucleotides that are useful for use in nucleic acid amplification reactions. These probes and oligonucleotides are modified to alter their sensitivity to primer-independent 5′ exonuclease activity of a thermostable DNA polymerase relative to its corresponding unmodified counterpart. Non-symmetric polymerase chain reaction (PCR) amplification and detection methods employing these labeled probes and unlabeled oligonucleotides are also described. Kits for nucleic acid amplification reactions including labeled probes and unlabeled oligonucleotides are also described.
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| Pcr-directed gene synthesis from large number of overlapping oligodeoxyribonucleotides | 20080182296 | 20080731 |
| The present invention provides methods of PCR-directed gene synthesis that may be used for all genes, including those with a high G+C content and/or a long sequence. The invention relates to methods of gene synthesis using overlapping oligonucleotides and polymerase chain reaction (PCR), wherein several PCR parameters, e.g., the concentration of overlapping oligonucleotides, the type of DNA polymerase used, and the number of PCR amplification cycles, are optimized. Additionally, the invention relates to oligonucleotide design that allows for increased protein expression of synthesized genes.
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| Heat and ph measurement for sequencing of dna | 20080166727 | 20080710 |
| The present method involves sequencing by synthesis in which a template strand having an attached primer is immobilized in a small volume reaction mixture. In one embodiment, the reaction mixture is in contact with a sensitive heat sensor, which detects the heat of reaction from incorporation of a complementary base (dNTP) in the presence of appropriate reagents (DNA polymerase, and polymerase reaction buffer). Alternatively, or in addition, a change in pH resulting from the incorporation of nucleotides in the DNA polymerase reaction is measured. A device is provided having delivery channels for appropriate reagents, including dNTPs, which may be delivered sequentially or in a mixture. Preferably, the dNTPs are added in a predetermined sequence, and the dNTP is incorporated or not depending on the template sequence.
... |
| Optical detection of analytes by use of semiconductor nanoparticles | 20080153085 | 20080626 |
| The present invention provides an optical method and device for the determination of an analyte in an assayed sample. The method and device of the invention are based on the use of semiconductor nanoparticles that carry a recognition agent. The detection is based on fluorescence resonance energy transfer (FRET) between the semiconductor nanoparticle donors, which are excited with electromagnetic radiation, and acceptors in the form of dye-labeled or semiconductor nanoparticle-labeled agents, that are immobilized to the recognition agent in the presence of the analyte and under assay conditions. The method and device of the present invention are especially useful in the determination of DNA or RNA analytes, DNA polymerase or telomerase analytes, cancer cells through telomerase activity and single-base mutations. In such cases the recognition agent... |
| Stabilized compositions of thermostable dna polymerase and anionic or zwitterionic detergent | 20080145910 | 20080619 |
| The present invention provides compositions, methods, and kits for protecting thermostable DNA polymerase during amplification reactions conducted at a temperature ranging from about 40° C. to greater than 100° C. The composition comprises a thermostable DNA polymerase and an anionic detergent or zwitterionic detergent.
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| Polymerase enzymes and reagents for enhanced nucleic acid sequencing | 20080108082 | 20080508 |
| Compositions that include DNA polymerases having increased residence times for nucleotide analogues, particularly modified recombinant Φ29-type DNA polymerases with such increased residence times, are provided. Methods of making the polymerases and of using the polymerases in sequencing and DNA amplification are also provided. Compositions including α-thiophosphate nucleotide analogues with four or more phosphate groups are described, as are methods for determining the sequence of nucleic acid molecules using such analogues.
... |
| Unstructured nucleic acid pcr primers and methods of using the same | 20080090235 | 20080417 |
| A polymerase chain reaction (PCR) mixture containing at least one unstructured nucleic acid primer pair is provided. In certain embodiments, the mixture may also contain: nucleotides, a DNA polymerase, and PCR reaction reagents, as well as a nucleic acid sample. The reaction mixture may be employed in, for example, a PCR reaction.
... |
| Rapid reverse transcription of rna | 20080085541 | 20080410 |
| Methods and kits for use in the practice of reverse transcriptase and for one-tube practice of reverse transcriptase and polymerase chain reaction are described. Use of an RNA template having a known conserved area of at least about 40 base pairs, a reverse transcription primer suitable for preparing DNA from the conserved area of said RNA template, and a reverse transcriptase having an elongation rate of at least 40 base pairs per second and a processivity of at least 75 base pairs allows for rapid reverse transcription of the conserved area. Use of PCR primers having 3′ ends which are separated by about 1 to about 120 base pairs, DNA polymerase having an elongation rate of at least 60 base pairs per second and a processivity... |
| Recombinase polymerase amplification | 20080076160 | 20080327 |
| This disclosure describe three related novel methods for Recombinase-Polymerase Amplification (RPA) of a target DNA that exploit the properties of the bacterial RecA and related proteins, to invade double-stranded DNA with single stranded homologous DNA permitting sequence specific priming of DNA polymerase reactions. The disclosed methods has the advantage of not requiring thermocycling or thermophilic enzymes. Further, the improved processivity of the disclosed methods allow amplification of DNA up to hundreds of megabases in length.
... |
| Zwitterionic detergents for the storage and use of dna polymerases | 20080064071 | 20080313 |
| The present invention provides methods, compositions, and kits for storing and enhancing the activity of thermostable polymerases. The methods comprise mixing a thermostable polymerase with at least one zwitterionic detergent or non-detergent surfactant. Compositions and kits for performing the process according to the invention are also provided.
... |
| Ambient temperature stable kits for molecular diagnostics | 20080050737 | 20080228 |
| A method for processing DNA polymerase and/or dNTPs for use in an amplification procedure, includes providing a solution mixture, the solution mixture including a DNA polymerase and/or dNTPs, a buffer solution and at least one stabilizing agent and hydration reducing the solution mixture. The solution mixture is hydration reduced at a temperature between 0 °C. and about 100 ° C.
... |
| Methods and compositions for amplification of dna | 20080044883 | 20080221 |
| The invention provides an Enzyme Blend comprising a DNA polymerase and a DNA repair enzyme. Methods and kits for amplification of DNA that is damaged, undamaged, or suspected of being damaged are also provided.
... |
| Methods for amplifying polymeric nucleic acids | 20080038724 | 20080214 |
| The invention provides compositions and methods for amplifying nucleic acid polymer sequences in a high complexity nucleic acid sample. The unique compositions of the invention include a primer set composed of a mixture of two types of primers for DNA synthesis. For extension in one direction, the primers all contain modifications that destroy their ability to serve as templates that can be copied by DNA polymerases. For extension in the opposite direction the set includes at least one primer that can serve as a template and be replicated by DNA polymerases throughout its length. The method can be carried out by mixing the nucleic acid polymer sequence of interest with the set of DNA synthesis primers in an amplification reaction mixture. The reaction mixture is then... |
| Pyrophosphorolysis activated polymerization (pap) | 20070298428 | 20071227 |
| A novel method of pyrophosphorolysis activated polymerization (PAP) has been developed. In PAP, pyrophosphorolysis and polymerization by DNA polymerase are coupled serially for each amplification by using an activatable oligonucleotide P* that has a non-extendible 3′-deoxynucleotide at its 3′ terminus. PAP can be applied for exponential amplification or for linear amplification. PAP can be applied to amplification of a rare allele in admixture with one or more wild-type alleles by using an activatable oligonucleotide P* that is an exact match at its 3′ end for the rare allele but has a mismatch at or near its 3′ terminus for the wild-type allele. PAP is inhibited by a mismatch in the 3′ specific sequence as far as 16 nucleotides away from the 3′ terminus. PAP can greatly... |
| Protein crystal comprising the processivity clamp factor of dna polymerase and a ligand, and its uses | 20070275898 | 20071129 |
| The present invention relates to a protein crystal comprising the processivity clamp factor of DNA polymerase and a peptide of about 3 to about 30 amino acids, in particular of about 16 amino acids, said peptide comprising all or part of the processivity clamp factor binding sequence of a processivity clamp factor interacting protein, such as prokaryotic Pol I, Pol II, Pol III, Pol IV, Pol V, MutS, ligase I, α subunit of DNA polymerase, UmuD or UmuD′, or eukaryotic pol ε, pol δ, pol η, pol τ, pol κ.
... |
| Diagnosis and prediction of parkinson's disease | 20070277251 | 20071129 |
| The present invention provides an in vitro method for diagnosis or prediction of Parkinson's disease in a subject, the method comprising testing for a mutation of a mitochondrial DNA polymerase POLG1 gene of the subject and, where the mutation is detected, diagnosing or predicting Parkinson's disease in the subject. Further, the present invention provides a diagnostic kit for diagnosis or prediction of Parkinson's disease in a subject.
... |
| Methods for genotyping | 20070269817 | 20071122 |
| The present invention provides for methods for discriminating between alleles at polymorphic positions in a genome. In general the methods employ allele specific extension of oligonucleotides that are complementary to one of the alleles at the 3′ end of the oligonucleotide. The allele specific oligonucleotides are resistant to proof reading activity from a polymerase and may be extended in an allele specific manner by a DNA polymerase with a functional 3′ to 5′ exonuclease activity. The allele specific oligonucleotides may be attached to a solid support such as a chip or a bead.
... |
| Dna polymerases | 20070269877 | 20071122 |
| The present invention relates to a variant archaeal DNA polymerase comprising a modified amino acid sequence of a wild-type amino acid sequence, wherein during DNA replication, the variant polymerase is adapted to mis-incorporate a greater number of incorrect bases than compared to a wild-type archaeal DNA polymerase. Such a variant DNA polymerase may be usefully employed in biological assay systems, such as PCR.
... |
| Hbv particles containing hbv-rna | 20070264646 | 20071115 |
| Hepatitis B virus (HBV) particles that contain HBV-RNA but do not contain HBV-DNA in a body fluid sample; A method of measuring the particles according to claim 1 by measuring HBV-RNA; A method of judging the therapeutic effect of the treatment of HBV infections by a drug having an effect of inhibiting RNA-dependent DNA polymerase, which method comprises performing the measurement on a body fluid of a patient who received said treatment; A method of predicting the risk of developing hepatic carcinoma or selecting risk groups of developing hepatic carcinoma by the above measurement.
... |
| Dna replication factors | 20070264660 | 20071115 |
| A DNA polymerase reaction system which provides high DNA polymerase activity even at a high temperature and at a high salt concentration. A DNA polymerase reaction system that is constructed from a DNA polymerase, a clamp, and a clamp loader without intein sequence, the DNA polymerase being from Pyrococcus horikoshii, a hyperthermophilic archaeon.
... |
| Single molecule drug discovery | 20070254279 | 20071101 |
| The present application discloses methods and apparatuses for single molecule drug screening, discovery and validation. These methods and apparatuses allow a user to detect rapidly, using observation of single molecules, whether and how a drug candidate interferes with a target enzyme involved in a particular disease pathway. The methods and apparatuses described herein utilize single molecule manipulation and detection technologies (e.g., optical or magnetic tweezers) to directly detect whether the characteristic dynamics, or “mechanical signature,” of the target enzyme-substrate interaction are substantially altered or modulated by a drug candidate. Furthermore, the methods and apparatuses are useful for analyzing the modulation of the mechanical signature in order to identify potential interference mechanisms of a drug candidate. In one aspect of the invention, the methods and apparatuses disclosed... |
| Helicase-dependent amplification of nucleic acids | 20070254304 | 20071101 |
| Methods and a kit are provided for selectively and exponentially amplifying nucleic acids and include the use of a helicase preparation and a DNA polymerase such that the amplification can be performed isothermally.
... |
| Random-primed reverse transcriptase - in vitro transcription method for rna amplification | 20070255053 | 20071101 |
| A random-primed reverse transcriptase-in vitro transcription method of linearly amplifying RNA is provided. According to the methods of the invention, source RNA (or other single-stranded nucleic acid), preferably, mRNA, is converted to double-stranded cDNA using two random primers, one of which comprises a RNA polymerase promoter sequence (“promoter-primer”), to yield a double-stranded cDNA that comprises a RNA polymerase promoter that is recognized by a RNA polymerase. Preferably, the primer for first-strand cDNA synthesis is a promoter-primer and the primer for second-strand cDNA synthesis is not a promoter-primer. The double-stranded cDNA is then transcribed into RNA by the RNA polymerase, optimally in the presence of a reverse transcriptase that is rendered incapable of RNA-dependent DNA polymerase activity during this transcription step. The subject methods produce linearly amplified... |
| Terminating substrates for dna polymerases | 20070248973 | 20071025 |
| This invention concerns novel terminating substrates for DNA polymerases. The substrates are nucleoside triphosphates labeled with lanthanide chelates.
... |
| Novel dna synthesis technology with 3'-beaded oligo dna and dna polymerase | 20070249024 | 20071025 |
| A method of synthesizing a desired DNA having a predetermined sequence, comprising the steps of (a) preparing, based on the desired DNA, a plurality of 3′-biotin-immobilized oligo DNA having a fixed length; (b) preparing a starting DNA which has a complementary sequence to a 3′ end of a first immobilized oligo DNA of the plurality of oligo DNA, and wherein the starting DNA has a length shorter than the length of the first immobilized oligo DNA; (c) annealing the starting DNA with the first immobilized oligo DNA, extending the starting DNA to complement the first immobilized oligo DNA, thereby making a newly extended DNA; (d) denaturing a first double strand DNA consisting of the newly extended DNA and the first immobilized oligo DNA; (e) collecting the... |
| Compositions and methods for reverse transcriptase-polymerase chain reaction (rt-pcr) of human b-retrovirus | 20070237716 | 20071011 |
| The invention can be summarized as follows. The present invention provides compositions, methods and kits useful for the amplification of nucleic acid molecules from human β-retrovirus by reverse transcriptase-polymerase chain reaction (RT-PCR). Specifically, the invention provides compositions and methods for the amplification of nucleic acid molecules in a one or two-step real time RT-PCR procedure using reverse transcriptase, DNA polymerase, or a combination of both enzymes. The invention provides for the rapid and efficient amplification, detection and quantification of human β-retrovirus nucleic acids.
... |
| Method for retaining methylation pattern in globally amplified dna | 20070238117 | 20071011 |
| A method for preserving information about cytosine methylation status in amplified nucleic acid molecules is disclosed. The method includes contacting a sample that contains nucleic acid molecules, such as nucleic acid molecules having or suspected of having methylated cytosines, with a modifying agent that converts the unmethylated cytosines to produce converted nucleic acid molecules. The converted nucleic acid molecule retains information about cytosine methylation. The method further involves contacting the sample with a DNA polymerase to amplify the converted nucleic acid molecules by multiple strand displacement amplification. The sample is not contacted with a nucleic acid ligase or an RNA polymerase. Also disclosed are methods for detecting cytosine methylation in a sample. Such methods include detecting the presence of the signature of cytosine methylation in a... |
| Methods of nucleic acid analysis by single molecule detection | 20070231808 | 20071004 |
| This invention provides a method of nucleic acid analysis that enables highly accurate and sensitive quantitation by counting the number of molecules among a plurality of types of genes without amplifying specific genes and that enable reduction of quantitation limits. This method comprises steps of: allowing a polynucleotide comprising a first region having a sequence complementary to the target gene at the 3′ end, a second region having a sequence complementary to the target gene at the 5′ end, and a third region corresponding to a detection probe to hybridize to the target gene; allowing the 3′ end of the first region hybridized to the target gene to ligate to the 5′ end of the second region so as to obtain a circularized polynucleotide; with the... |
| Method for detecting the presence of water born pathogens and indicator microorganism | 20070218468 | 20070920 |
| The present invention relates to a method for detecting the presence of water born pathogens and indicator microorganism including bacteria from water sample by selecting the target gene carried in template DNA by amplifying the target DNA using specific primers with biotinylated tag consist of all or a substantial part of 5′-CTGATCGAATGGCTGCCAGGCTCC-3′ and 5′-CAACCAGACGATAGTTATCACGCA-3′ and taq DNA polymerase to get desired biotinylated tagged probe followed by hybridization of biotinylated tagged probe with target gene in template DNA followed by enzyme coupled reaction.
... |
| Methods for dna amplification and sequencing | 20070212760 | 20070913 |
| The osmoprotectants proline, 2-methyl-4-carboxy-3,4,5,6-tetrahydropyrimidine (“THP(B)”, and 2-methyl-4-carboxy-5-hydroxy-3,4,5,6,-tetrahydropyrimidine (“THP(A)”) are capable of increasing the thermal stability of DNA polymerases at elevated temperatures. THP(B) is further effective in lowering the melting temperature of double-stranded DNA. Proline, THP(A) and THP(B) are thus useful in procedures involving melting of double-stranded DNA and/or polymerase-mediated DNA synthesis, such as in primer extension, in PCR (polymerase chain reaction) amplification and in DNA sequencing.
... |
| Helicase-dependent amplification of nucleic acids | 20070207495 | 20070906 |
| Methods and a kit are provided for selectively and exponentially amplifying nucleic acids and include the use of a helicase preparation and a DNA polymerase such that the amplification can be performed isothermally.
... |
| Method for controlled induction of somatic mutations and use thereof in proteomics | 20070196368 | 20070823 |
| The invention concerns a method for inducing somatic mutations in vitro or ex vivo, comprising (1): adding to the cells to be mutated, in culture conditions and in a medium suited to said cells, of at least a combination of anti-IgM and anti-CD19 and/or anti-DC21 antibodies, while (2) said anti-IgM antibodies are biotinylated and are subjected to specific aggregation by streptavidin coupled with a support. Advantageously, the mutations are controlled and/or adjusted by inhibiting and reactivating and/or stimulating DNA polymerase pol iota. The invention is in particular applicable to rapid and controlled induction of mutations on the BL2 Burkitt's lymphoma.
... |
| Method for long range allele-specific pcr | 20070184457 | 20070809 |
| The present invention is directed to a method and kit for determining the molecular haplotype of a gene in a diploid DNA sample. The method discriminates between two haplotypes on the basis of a difference of one or more nucleotides using allele-specific PCR amplification in combination with long range PCR, using a DNA polymerase enzyme having 3′→5′ exonuclease activity, using annealing temperature conditions sufficiently greater than the predicted annealing temperature (Tm) to effect selective hybridization and extension of an allele-specific extension primer to the target allele relative to the variant allele. The present invention is particularly useful, for example, to determine the haplotype of a gene having multi-allelic genetic loci separated by a distance in which accurate PCR amplification of a single fragment containing the multiple... |
| Apparatus and method for solid-phase kinetic analysis of templated elongation reactions | 20070184462 | 20070809 |
| Disclosed is an elongation reaction system that includes a membrane compatible with binding an elongation complex and an elongation complex compatible with binding the membrane. The elongation complex includes the biological template (e.g., DNA or RNA), a polymerizing agent (e.g., RNA polymerase, DNA polymerase, or a ribosome), and a primer transcript or polypeptide. Further, disclosed is an apparatus and method for solid-phase kinetic analysis of templated elongation reactions.
... |
| Methods for detecting and localizing dna mutations by microarray | 20070184468 | 20070809 |
| This disclosure provides methods for detecting and localizing DNA mutations by DNA microarray. In various embodiments, the described methods include use of restriction endonuclease(s) and/or mismatch-recognition nuclease(s) to detect and/or localize mutations. In one representative method, reference and target DNA are digested using one or more restriction endonucleases, resultant DNA strands are labeled (e.g., using a DNA polymerase), and the labeled mixture of DNAs is hybridized to a microarray. In another representative method, reference and target DNA are denatured and annealed to form a mixture containing heteroduplex DNA, one or more mismatch-recognition nuclease(s) are used to nick or cleave at least a portion of the heteroduplex DNA, resultant DNA strands are labeled (e.g., using a DNA polymerase) and the labeled mixture of DNAs is hybridized to... |
| Methods of random mutagenesis and methods of modifying nucleic acids using translesion dna polymerases | 20070184482 | 20070809 |
| The invention is related generally to methods of amplifying or synthesizing or producing nucleic acid molecules using Translesion DNA polymerases. In particular, the invention relates to methods of introducing a random mutation into a nucleic acid and encoded polypeptide using Translesion DNA polymerases. The invention also relates to methods of introducing a modified nucleotide into a nucleic acid using Translesion DNA polymerases. The invention also relates to mutagenized and modified nucleic acid molecules and proteins produced by these methods, and to fragments or derivatives thereof. The invention also relates to vectors and host cells comprising mutagenized nucleic acid molecules, fragments, or derivatives. The invention also relates to the use of mutagenized nucleic acid molecules to produce desired polypeptides and uses of modified nucleic acid molecules to... |
| Compositions and methods for synthesizing nucleic acids | 20070178491 | 20070802 |
| Disclosed are compositions preferably for use in nucleic acid synthesis that include one or more anti-reverse transcriptase (RT) antibodies and/or one or more anti-DNA polymerase (DNAP) antibodies and/or single strand binding proteins (SSBs). Some of the disclosed compositions include one or more anti-DNAP antibodies and/or one or more anti-RT antibodies and one or more SSBs. Other disclosed compositions include two or more SSBs. The disclosed nucleic acid synthesis compositions also can include one or more DNAPs, one or more RTs, one or more nucleotides, one or more primers, and/or one or more templates. Also disclosed are methods for using such compositions in nucleic acid synthesis, amplification and sequencing, where various combinations of anti-RT antibodies, anti-DNAP antibodies and/or SSBs can improve the yield and/or homogeneity of primer... |
| Thermophilic dna polymerases from thermoactinomyces vulgaris | 20070172879 | 20070726 |
| The present invention provides compositions comprising thermostable DNA polymerases derived from hyperthermophilic eubacteria. In particular, the present invention comprises thermostable DNA polymerases from the hyperthermophilic eubacterial species Thermoactinomyces vulgaris. The present invention also provides methods for utilizing naturally-occurring and non-naturally-occurring forms of T. vulgaris DNA polymerase in sequencing, reverse transcription, and amplification reactions.
... |
| Methods and compositions for mutation analysis | 20070161010 | 20070712 |
| In one aspect, a method for DNA mutation detection including the steps of PCR amplification using preferably Pho DNA polymerase, hybridization, and analysis by denaturing high performance liquid chromatography (DHPLC), the method preferably utilizing a PCR buffer and other solutions that are compatible with DHPLC analysis. In other aspects, compositions and kits including a proofreading DNA polymerase, preferably Pho DNA polymerase, and a DHPLC compatible PCR buffer are provided.
... |
| Method for preparing active nanoarchaeum equitans dna polymerase and the active dna polymerase prepared by the method | 20070154913 | 20070705 |
| Disclosed are a method of preparing an active Nanoarchaeum equitans B-type DNA polymerase (Neq DNA polymerase), an active Neq DNA polymerase prepared according to the method, and a polymerase chain reaction (PCR) using the active Neq DNA polymerase. The active Neq DNA polymerase may be used in various nucleic acid polymerization reactions, such as PCR.
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| Dna polymerase fusions and uses thereof | 20070148671 | 20070628 |
| The present invention discloses methods of using DNA polymerase fusions at high pH in PCR, DNA sequencing and mutagenesis protocols.
... |
| Dna binding protein-polymerase chimeras | 20070141591 | 20070621 |
| The invention relates to compositions and methods directed to chimeric DNA polymerases, which comprise a mutated DNA binding polypeptide domain and a mutated or wild-type DNA polymerase polypeptide domain.
... |
| Oligonucleotides for detection of leishmaniasis and methods thereof | 20070134671 | 20070614 |
| The present invention relates to a novel oligonucleotide primers having SEQ ID NO: 1, SEQ ID No: 2, SEQ ID NO: 3 and SEQ ID NO: 4 for amplification of the kinesin-related gene of Leishmania species. The invention also provides a method for detecting and differentiating visceral leishmaniasis (VL) and post kala-azar-dermal leishmaniasis (PKDL) causing strains of Leishmania donovani in a sample, comprising isolating DNA from a sample; amplifying the target region from the DNA using novel oligonucleotide primers and heat stable DNA polymerase to obtain amplified fragments; separating the amplified fragments and analyzing the fragments to detect and differentiate VL and PKDL causing strains of Leishmania donovani based on the banding pattern of the amplified fragments. In addition, the invention provides a diagnostic kit for... |
| Dna polymerase blends and mutant dna polymerases | 20060292578 | 20061228 |
| A thermostable DNA polymerase composition comprising at least two DNA polymerases, one of which is substantially reduced in 5′-exonuclease activity and one of which has 5′-exonuclease activity. This polymerase may be used in methods including, but not limited to, nucleic acid synthesis, DNA sequencing, nucleic acid amplification and cDNA synthesis,
... |
| Methods for purifying dna polymerases | 20060286562 | 20061221 |
| The present invention provides methods and kits for obtaining substantially pure DNA polymerases. The methods comprise fractionating preparations comprising at east one DNA polymerase using Poly U Sepharose chromatography and obtaining substantially pure DNA polymerase. The present invention also provides compositions comprising substantially pure archaebacterial DNA polymerase obtained by fractionation using Poly U Sepharose chromatography resin.
... |
| Polymerases | 20060281109 | 20061214 |
| Modified DNA polymerases have an affinity for DNA such that the polymerase has an ability to incorporate one or more nucleotides into a plurality of separate DNA templates in each reaction cycle. The polymerases are capable of forming an increased number of productive polymerase-DNA complexes in each reaction cycle. The modified polymerases may be used in a number of DNA sequencing applications, especially in the context of clustered arrays.
... |
| Compositions and methods for polynucleotide amplification and detection | 20060269932 | 20061130 |
| Described are characteristics rendering DNA polymerases well suited for single target and multiplex genetic profiling methods, and polymerases having these characteristics. Also described are methods of single target and multiplex analyses using polymerases having the described characteristics.
... |
| Methods of copying the methylation pattern of dna during isothermal amplification and microarrays | 20060257905 | 20061116 |
| A method for copying the methylation patterns of molecules of genomic DNA (MGD) during isothermal amplification of the MGD comprising obtaining MGD, copying the methylation patterns of the MGD using a DNA methylation-maintenance enzyme, while isothermally amplifying the MGD using a DNA polymerase with strand displacement activity, under conditions that simultaneously promote activity of the DNA methylation-maintenance enzyme and the DNA polymerase; a method for copying the methylation patterns in double-stranded DNA molecules during isothermal amplification of the DNA molecules comprising obtaining DNA molecules, contacting the DNA molecules with transposable elements and an enzyme, which can randomly insert the transposable elements into the DNA molecules, copying the methylation patterns of the DNA molecules using a DNA methylation-maintenance enzyme, while isothermally amplifying the DNA molecules using a... |
| Antibacterial pyrazole carboxylic acid hydrazides | 20060258729 | 20061116 |
| Methods and compositions comprising pyrazole carboxylic acid hydrazide compounds are described that are useful for treating bacterial infections, inhibiting bacterial growth, and inhibiting the activity of bacterial DNA polymerase III.
... |
| Methods and compositions for detecting telomerase activity | 20060246441 | 20061102 |
| A method for determining telomerase activity using primer extension followed with real time PCR quantification is disclosed. The method of the present invention provides a rapid, sensitive and accurate measurement for telomerase activity in a biological sample. In one embodiment, the method includes the steps of: adding the biological sample to a reaction tube containing a first reaction mixture having a first primer and nucleoside triphosphates, a second reaction mixture having a second primer and a DNA polymerase, and a wax layer that separates the first reaction mixture from the second reaction mixture; incubating the biological sample with the first reaction mixture; admixing the extension product with the second reaction mixture; amplifying and quantifying the extension product using real-time PCR and a control template. In another... |
| Compositions for dna amplification, synthesis, and mutagenesis | 20060246462 | 20061102 |
| This invention provides compositions comprising a thermostable non-proofreading DNA polymerase, a thermostable proofreading DNA polymerase, and a factor that substantially inhibits the incorporation of undesired nucleotides or analogs thereof into a DNA polymer. The compositions may further comprise a buffer that enhances a polymerization reaction involving DNA polymerases. The invention also provides various methods of amplifying, synthesizing, or mutagenizing nucleic acids of interest using these novel compositions. Kits that comprise the compositions are also provided for amplifying, synthesizing, and mutagenizing nucleic acids.
... |
| Method of reverse transcription | 20060240468 | 20061026 |
| The present invention relates to reverse transcription of RNA, and in particular to reverse transcription by thermostable DNA polymerases. Thermoactinomyces vulgaris and Bacillus stearothermophilus possess reverse transcriptase activity in the presence of magnesium or manganese ions. Methods, compositions, and kits for reverse transcription and RT-PCR are also provided.
... |
| Helix-hairpin-helix motifs to manipulate properties of dna processing enzymes | 20060234227 | 20061019 |
| The utility of Topo V's HhH motifs for modulating DNA binding properties of a DNA processing enzyme, such as Taq DNA polymerase or PfuDNA polymerase, is demonstrated. In one embodiment, an amino acid residue comprising one or more HhH domains derived from Topoisomerase V, linked to either the NH2-terminus or COOH-terminus of a Taq polymerase fragment significantly broadens the processivity and/or salt concentration range of polymerase activity. The specific activities of the chimeric polymerases are not affected by added HhH motifs. Depending on the type of the construct, the thermal stability of chimeric polymerases increases or remains the same as that of Taq DNA polymerase or its Stoffel fragment. This invention further provides a method of increasing the salt tolerance of Taq polymerases. The methods of... |
| Multiplex polynucleotide synthesis | 20060234264 | 20061019 |
| The invention provides a method of synthesizing complex mixtures of long polynucleotides by separately synthesizing and assembling shorter component oligonucleotides. In one aspect, pairs of oligonucleotides that form components of such polynucleotides are synthesized on one or more microarrays, or other large-scale parallel solid phase synthesis platforms, after which they are released. Members of each pair contain unique complementary barcode sequences that are used match-up pairs in a hybridization reaction to form duplexes. Such duplexes are then extended with a DNA polymerase and the resulting extension product is amplified to form an amplicon. The amplicon may be either used directly as the desired polynucleotide, or it may undergo further processing, such as capture on solid phase supports and/or additional enzymatic or chemical processing, to produce a... |
| Mutant dna polymerases and methods of use | 20060223067 | 20061005 |
| The present invention provides mutant DNA polymerases, polynucleotides encoding the polymerases, cassettes and vectors including such polynucleotides, and cells containing the polymerases, polynucleotides, cassettes, and/or vectors of the invention. The present invention also provides methods for synthesizing polynucleotides and kits including a DNA polymerase of the invention.
... |
| Polypeptides having nucleic acid binding activity | 20060223157 | 20061005 |
| Polypeptides having nucleic acid binding activity are provided. Methods of stabilizing a nucleic acid duplex are provided. Methods of promoting the annealing of complementary nucleic acid strands are provided. Methods of increasing the processivity of a DNA polymerase are provided. Methods of enhancing the activity of a nucleic acid modification enzyme are provided. Fusion proteins are provided. Methods of using fusion proteins are provided. Kits are provided.
... |
| One component and two component dna pol lll replicases and uses thereof | 20060211003 | 20060921 |
| The invention provides one-component and two-component DNA polymerases, as well as kits comprising the same, and methods of using the same for nucleic amplification and nucleic acid sequencing.
... |
| Mouse model for aging | 20060212954 | 20060921 |
| A mouse model for mammalian aging is disclosed. In one embodiment, the invention comprises a mouse having a genomic mutation in the exonuclease domain II (ExoII) of a mitochondrial DNA polymerase gamma (PolG) gene, wherein the mutation leads to high levels of mutations in polymerase mtDNA.
... |
| Methods for synthesizing polynucleotides using a single primer | 20060204973 | 20060914 |
| An objective of the present invention is to provide methods for synthesizing polynucleotides comprising an unknown nucleotide sequence in a region containing their 5′-end. By using a single gene-specific primer, nucleotide sequences complementary to each other are added respectively at the 5′- and 3′-ends of a desired polynucleotide, to synthesize a desired polynucleotide. Requisite nucleotide sequences can be easily added by incubating with a primer, double-stranded DNA, and template-dependent DNA polymerase. By applying the present invention to 5′ RACE, genes expressed in a low-level, which cannot be obtained by known methods, have been efficiently obtained.
... |
| Incorporation of modified nucleotides by archaeon dna polymerases and related methods | 20060199214 | 20060907 |
| The present invention is directed toward improving the efficiency of chain terminator incorporation by Family B archaeon DNA polymerases. Previously, the low efficiency of ddNTP, and more especially dye-labeled ddNTP, incorporation has limited the usefulness of this group of DNA polymerases in protocols requiring chain terminator incorporation.
... |
| Viral agents | 20060188522 | 20060824 |
| A viral DNA construct, and virus encoded thereby, is provided having one or more tumour specific transcription factor binding sites in place of one or more wild type transcription factor binding sites operatively positioned in the promoter region which controls expression of early genes responsible for viral nucleic acid replication. Preferred constructs place the tumour specific transcription factor binding sites in operative relation to DNA polymerase, DNA terminal protein and/or DNA binding protein. Compositions and constructs contained therein are provided, particularly for use in therapy. Methods of treating patients for neoplasms are also provided.
... |
| 5-methylcytosine detection, compositions and methods therefor | 20060172309 | 20060803 |
| Compositions and methods for detecting 5-methylcytosine in a nucleic acid are disclosed. A 5-methylcytosine discriminator, which is a deoxyribonucleosidetriphosphate comprising a cytosine-pairing moiety such as a guanosine and a moiety which hinders hydrogen bonding between the cytosine-pairing moiety and a 5-methylcytosine is described. The discriminator is able to base pair with a cytosine but not a 5-methylcytosine. A 5-methylcytosine comprised by a target nucleotide can be detected in a reaction using a DNA polymerase and a primer hybridized immediately adjacent to the target nucleotide. In the reaction, pyrophosphate released upon incorporation of a dNTP complementary to a target nucleotide is detected. Lack of incorporation of the discriminator, but incorporation of a dGTP, can indicate that the target nucleotide is a 5-methylcytosine.
... |
| Combinations of inhibitors of reverse transcriptase and inhibitors of virus-encoded dna polymerase | 20060172997 | 20060803 |
| A method for treating viral diseases which are caused by DNA viruses is described comprising administering an effective amount of at least one inhibitor of reverse transcriptase (RTI) in combination with at least one inhibitor of viral DNA polymerase, wherein the at least one RTI and the at least one DNA polymerase inhibitor are present in the form of separate compounds.
... |
| Hbv drug resistance drug resistance detection methods | 20060165725 | 20060727 |
| New polymorphisms in the nucleic acid sequences of the DNA polymerase/reverse transcriptase open reading frame and viral surface antigen open reading frame of the hepatitis B virus are reported. In particular, the present invention relates to the mutation YMDD YSDD in the HBV reverse transcriptase domain and to the W196V mutation in the small HBV viral surface antigen. Said polymorphisms are affecting the detection of drug resistance mutations by genotypic methods and diagnostic kits based thereon. The present invention relates to methods and diagnostic kits for detection of a HBV virus comprising said nucleic acid polymorphisms. In particular, those methods utilizing oligonucleotides capable of hybridizing to said HBV nucleic acid polymorphisms are envisaged.
... |
| Helicase-dependent amplification of nucleic acids | 20060154286 | 20060713 |
| Methods and a kit are provided for selectively and exponentially amplifying nucleic acids and include the use of a single strand helicase preparation or a thermostable helicase in the absence of a single strand binding protein and a DNA polymerase such that the amplification can be performed isothermally.
... |
| Phi 15 dna polymerase | 20060147967 | 20060706 |
| A purified recombinant DNA polymerase having high processivity and strand-displacement activity. An isolated nucleic acid that encodes the bacteriophage DNA polymerase. A kit and method for amplifying DNA is also disclosed.
... |
| Novel polymerase compositions and uses thereof | 20060088822 | 20060427 |
| The subject invention provides novel compositions containing a mixture of (a) an enzyme that possesses substantial 3′-5′ exonuclease activity (b) a DNA polymerase with less 3′-5′ exonuclease activity than the enzyme with substantial 3′-5′ exonuclease activity. Preferably, the DNA polymerase for inclusion in the compositions are DNA polymerases that substantially lack 3′-5′ exonuclease activity. A preferred embodiment of the invention is a composition comprising the Taq DNA polymerase (isolated from Thermus aquaticus) and the Pfu DNA polymerase (isolated from Pyrococcus furiosus). Another aspect of the invention is to provide methods for synthesizing polynucleotides, typically DNA, using compositions comprising an enzyme that possesses substantial 3′-5′ exonuclease activity and a DNA polymerase with less 3′-5′ exonuclease activity than the enzymes possessing substantial 3′-5′ exonuclease activity, preferably a DNA... |
| Dna polymerases having improved labeled nucleotide incorporation properties | 20060088879 | 20060427 |
| The present invention relates to mutant DNA polymerases that exhibit reduced discrimination against labeled nucleotides into polynucleotides. The DNA polymerases of the invention have at least one mutation in the nucleotide label interaction region of the enzyme such the mutation results in reduced discrimination against labeled nucleotides. The nucleotide label interaction regions is located at portions of the O-helix, (ii) the K helix, and (iii) the inter O—P helical loop of Taq DNA polymerase or analogous positions in other DNA polymerases. In addition to providing novel mutant DNA polymerases, the invention also provides polynucleotides encoding the subject mutant DNA polymerases. The polynucleotides provided may comprise expression vectors for the recombinant production of the mutant polymerases. The invention also provide host cells containing the subject polynucleotides. The... |
| Thermostable enzyme promoting the fidelity of thermostable dna polymerases-for improvement of nucleic acid synthesis and amplification in vitro | 20060078928 | 20060413 |
| A purified thermostable enzyme is derived form the thermophilic archaebacterium Archaeoglobus fulgidus. The enzyme can be native or recombinant, is stable under PCR conditions and exhibits double strand specific exonuclease activity. It is a 3′-5′ exonuclease and cleaves to produce 5′-mononucleotides. Thermostable exonucleases are useful in many recombinant DNA techniques, in combination with a thermostable DNA polymerase like Taq especially for nucleic acid amplification by the polymerase chain reaction (PCR).
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| Methods for amplifying and analyzing nucleic acids | 20060073511 | 20060406 |
| The present invention provides methods for reducing the complexity of a nucleic acid sample to interrogate a collection of target sequences. Complexity reduction can be accomplished by annealing one or more target-specific primers to a nucleic acid sample containing genomic DNA and elongating the primers using a DNA polymerase with a high processivity rate. Labeled nucleotides or a labeled primer may be incorporated into the extension products and the labeled extension products may be separated from the unlabeled nucleic acid by affinity purification. The enriched sample may be further amplified using a target specific or non-specific amplification method. The invention further provides for analysis of the above sample to interrogate sequences of interest such as polymorphisms and to detect translocations and map translocation breakpoints. The amplified... |
