JP4960250B2 - 酵素によるdna合成の特異性を増大するための方法 - Google Patents
酵素によるdna合成の特異性を増大するための方法 Download PDFInfo
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- JP4960250B2 JP4960250B2 JP2007542117A JP2007542117A JP4960250B2 JP 4960250 B2 JP4960250 B2 JP 4960250B2 JP 2007542117 A JP2007542117 A JP 2007542117A JP 2007542117 A JP2007542117 A JP 2007542117A JP 4960250 B2 JP4960250 B2 JP 4960250B2
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- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
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- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6813—Hybridisation assays
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- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/686—Polymerase chain reaction [PCR]
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- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6869—Methods for sequencing
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- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
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- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Pyridine Compounds (AREA)
Description
R1、R2、R3、R4、R5は同一又は異なるものであって良く、-H、-OH、-F、-Cl、-Br、-CF3、-CCl3、-CBr3、-NH2、-NO2、-CN、-CONH2、-CHO、-CO2H、-SH、-SO3H、-OPO3H2、及びR7基からなる群から独立に選択され、
R6は、-O-、-OPO3H2、-CF3、-CCl3、-CBr3、-CN、-CONH2、-CHO、-CO2H、及びR7基からなる群から選択され、
各々のR7は、C1-6アルキル、C2-6アルケニル、C2-6アルキニル、又は5から8員環のアリール若しくはヘテロアリール環(ヘテロアリール環である場合は、1又は2つの窒素、酸素、又は硫黄原子あるいはそれらの混合物を含み、残部が炭素であって良い)から独立に選択され、R7は、-OH、-F、-Cl、-Br、-I、-CF3、-CCl3、-CBr3、-NH-NH2、-NH2、-NO、-NO2、-CN、-CONH2、-CHO、-CO2H、-SH、-SO3H、-OPO3H2、又はそれらの混合物から選択される1から4つの基によって任意に置換され、更に任意に、R7は、-NH-NH-、-NN-、-NH-、-CO-、-S-S-基を含んで良い。
正に荷電したピリジニウム基を分子内に含有する化合物(ピリジン-N-オキシド及び3-(1-ピリジニオ)-1-プロパンスルホネート)を添加することによるPCRの特異性及び収量の改善。
ベタイン、テトラメチルアンモニウムクロリド(TMA-Cl)、ジメチルスルホキシド(DMSO)、及び3-(1-ピリジニオ)-1-プロパンスルホネートのPCRの特異性を改善する能力の比較。
トリメチルアミン-N-オキシド(米国特許第6,270,962号)及びピリジン-N-オキシド(本発明)のPCRの特異性及び収量を増大する能力の比較。
Claims (14)
- DNAポリメラーゼを含有する反応混合物にピリジン-N-オキシドまたは3-(1-ピリジニオ)-1-プロパンスルホネートを含ませることによって、DNAポリメラーゼの反応を実施する方法。
- 前記DNAポリメラーゼの反応がPCRである、請求項1に記載の方法。
- 前記DNAポリメラーゼの反応がチェーンターミネーションDNA配列決定の反応である、請求項1に記載の方法。
- 前記DNAポリメラーゼの反応がプライマー伸長反応である、請求項1に記載の方法。
- 前記DNAポリメラーゼの反応が逆転写反応である、請求項1に記載の方法。
- 前記DNAポリメラーゼの反応がニックトランスレーション反応である、請求項1に記載の方法。
- 前記反応混合物がGCリッチのトリヌクレオチドリピートを有するDNA分子を含む、請求項1に記載の方法。
- 別々の容器に
a)DNAポリメラーゼの反応のための成分;及び
b)ピリジン-N-オキシドまたは3-(1-ピリジニオ)-1-プロパンスルホネートを含有する容器
を含む、DNAポリメラーゼの反応を実施するためのキット。 - DNAポリメラーゼ酵素を更に含む、請求項8に記載のキット。
- 前記DNAポリメラーゼ酵素が、E. coli DNAポリメラーゼI、クレノー酵素、T4 DNAポリメラーゼ、及びT7 DNAポリメラーゼからなる群から選択される、請求項9に記載のキット。
- 前記DNAポリメラーゼが逆転写酵素である、請求項9に記載のキット。
- 前記逆転写酵素がAMV及びM-MuLVからなる群から選択される、請求項11に記載のキット。
- 前記DNAポリメラーゼが熱安定性DNAポリメラーゼである、請求項9に記載のキット。
- 前記熱安定性DNAポリメラーゼが、Taq DNAポリメラーゼ;TaqポリメラーゼのN-末端欠失体、Tth DNAポリメラーゼ、Bacillus caldotenax DNAポリメラーゼ;Thermus flavus DNAポリメラーゼ;Bacillus stearothermophilus DNAポリメラーゼ;並びに古細菌DNAポリメラーゼからなる群から選択される、請求項13に記載のキット。
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GB0426303A GB2420560A (en) | 2004-11-30 | 2004-11-30 | A method for increasing the specificity of enzymatic DNA synthesis |
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PCT/GB2005/004537 WO2006059075A1 (en) | 2004-11-30 | 2005-11-25 | A method for increasing the specificity of enzymatic dna synthesis |
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EP2069487B1 (en) * | 2006-07-25 | 2014-03-19 | Agilent Technologies, Inc. | Zwitterionic detergents for the storage and use of dna polymerases |
JP5600880B2 (ja) * | 2009-03-03 | 2014-10-08 | 東洋紡株式会社 | 改良された増幅試薬 |
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US4683195A (en) | 1986-01-30 | 1987-07-28 | Cetus Corporation | Process for amplifying, detecting, and/or-cloning nucleic acid sequences |
US4683202A (en) | 1985-03-28 | 1987-07-28 | Cetus Corporation | Process for amplifying nucleic acid sequences |
US4889818A (en) | 1986-08-22 | 1989-12-26 | Cetus Corporation | Purified thermostable enzyme |
DE69531487T2 (de) * | 1994-01-31 | 2004-06-17 | The Regents Of The University Of California, Oakland | Methode zur eliminierung von sequenzierartefakten |
DE4411588C1 (de) * | 1994-03-30 | 1995-09-28 | Deutsches Rheuma Forschungszen | Puffer für DNA- und RNA-Polymerase-Reaktionen |
US6270962B1 (en) * | 1995-01-30 | 2001-08-07 | The Regents Of The University Of California | Methods for the elimination of DNA sequencing artifacts |
US6787305B1 (en) * | 1998-03-13 | 2004-09-07 | Invitrogen Corporation | Compositions and methods for enhanced synthesis of nucleic acid molecules |
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JP2011254833A (ja) | 2011-12-22 |
JP2008521395A (ja) | 2008-06-26 |
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