JPH05507000A - ヌクレオシド−5’−o−(1−チオトリホスフェート)を用いる改良された核酸シーケンス分析 - Google Patents
ヌクレオシド−5’−o−(1−チオトリホスフェート)を用いる改良された核酸シーケンス分析Info
- Publication number
- JPH05507000A JPH05507000A JP91517034A JP51703491A JPH05507000A JP H05507000 A JPH05507000 A JP H05507000A JP 91517034 A JP91517034 A JP 91517034A JP 51703491 A JP51703491 A JP 51703491A JP H05507000 A JPH05507000 A JP H05507000A
- Authority
- JP
- Japan
- Prior art keywords
- dye
- nucleic acid
- dideoxy
- dyes
- bonded
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- 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
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S435/00—Chemistry: molecular biology and microbiology
- Y10S435/968—High energy substrates, e.g. fluorescent, chemiluminescent, radioactive
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S436/00—Chemistry: analytical and immunological testing
- Y10S436/80—Fluorescent dyes, e.g. rhodamine
Abstract
Description
Claims (16)
- 1.標的核酸のヌクレオチドシーケンスを決定する方法が:鋳型核酸を用意し、 この鋳型核酸は標的核酸を含有しており;鋳型核酸のためのオリゴヌクレオチド プライマーを用意し;オリゴヌクレオチドプライマーに鋳型核酸に対してハイブ リッドを形成し; 核酸ポリメラーゼを用いてヌクレオシド−5′−O−(1−チオトリホスフェー ト)前駆体および少なくとも1種の鎖終結ヌクレオチドを含有する反応混合物に おいてオリゴヌクレオチドプライマーを伸長し、同じ大きさの各DNAフラグメ ントが同じ鎖終結ヌクレオチドを用いて終結するようにDNAフラグメント個体 群のネストシリーズを形成し; DNAフラグメント個体群を大きさによって分離し;そして 各DNAフラグメント個体群と結合した鎖終結ヌクレオチドを同定する各工程か ら成る方法。
- 2.前記分離の工程がゲル電気泳動によって前記DNAフラグメント個体群を分 離し、各バンドが同じ大きさのDNAフラグメントに対応するようにDNAフラ グメント個体群のバンドを形成する請求項1記載の方法。
- 3.前記少なくとも1種の鎖終結ヌクレオチドが第1の染料で標識化されたジデ オキシアデノシン、第2の染料で標識化されたジデオキシシチジン、第3の染料 で標識化されたジデオキシグアノシン、および第4の染料で標識化されたジデオ キシチミジンから成り、第1、第2、第3、および第4の染料が一方が他方に関 してスペクトルにより分割でき、そして前記同定する工程は第1、第2、第3、 および第4の染料が蛍光を発生する原因となるようにDNAフラグメントの前記 バンドを発光し、第1、第2、第3、および第4の染料の蛍光および/または吸 着特性を測定する各工程を含む請求項2記載の方法。
- 4.前記核酸ポリメラーゼがシーケナーゼ(Sequenase(登録商標)) である請求項3記載の方法。
- 5.前記ジデオキシアデノシンが2′,3′−ジデオキシ−7−デアザアデノシ ンであり、前記第1の染料が連結基によってその7炭素原子に結合しており;前 記ジデオキシシチジンが2′,3′−ジデオキシシチジンであり、前記第2の染 料が連結基によってその5炭素原子に結合しており;前記ジデオキシグアノシン が2′,3′−ジデオキシ−7−デアザグアノシンおよび2′,3′−ジデオキ シ−7−デアザイノシンから成る群から選ばれ、前記第3の染料が連結基によっ てその7炭素原子に結合し;そして前記ジデオキシチミジンが2′,3′−ジデ オキシウリジンであり、前記第4の染料が連結基によってその5炭素原子に結合 している請求項4記載の方法。
- 6.前記第1、第2、第3、および第4の染料がローダミン染料およびフルオレ セイン染料から成る群から選ばれる請求項5記載の方法。
- 7.前記連結基が3−カルボキシアミノ−1−プロビニルであり、前記2′,3 ′−ジデオキシ−7−デアザアデノシン、および2′,3′−ジデオキシ−7− デアザグアノシンおよび2′,3′−ジデオキシ−7−デアザイノシンから成る 群から選ばれる前記ジデオキシグアノシンの前記7炭素原子を、それぞれ、前記 第1および第3の染料の5または6炭素原子に結合し、そして前記ジデオキシシ チジンおよび前記ジデオキシチミジンの前記5炭素原子をそれぞれ、前記第2お よび第4の染料の5または6炭素原子に結合する請求項6記載の方法。
- 8.前記第1、第2、第3、および第4の染料がローダミンX、テトラメチルロ ーダミン、ローダミン110、およびローダミン6Gから選ばれる請求項7記載 の方法。
- 9.前記2′,3′−ジデオキシ−7−デアザアデノシンをローダミン6Gの前 記5炭素原子に結合し、そして前記2′,3′−ジデオキシシチジンをローダミ ンXの前記6炭素原子に結合し、2′,3′−ジデオキシ−7−デアザグアノシ ンおよび2′,3′−ジデオキシ−7−デアザイノシンから成る群から選ばれる 前記ジデオキシグアノシンをローダミン110の前記5炭素原子に結合し、前記 2′,3′−ジデオキシウリジンをテトラメチルローダミンの前記6炭素原子に 結合する請求項8記載の方法。
- 10.前記第1、第2、第3、第4の染料が5−(および6−)カルボキシフル オレセイン、2′,7′−ジクロロ−5−(および6−)カルボキシフルオレセ イン、2′,7′−ジメトキシ−4′,5′−ジクロロ−5−(および6−)カ ルボキシ−4,7−ジクロロフルオレセイン、および1′,2′,7′,8′− ジベンゾ−5−(および6−)カルボキシ−4,7−ジクロロフルオレセインか ら成る群から選ばれる請求項7記載の方法。
- 11.前記2′,3′−ジデオキシ−7−デアザアデノシンが2′,7′−ジメ トキシ−4′,5′−ジクロロ−4,7−ジクロロフルオレセインの前記5炭素 原子に結合し、前記2′,3′−ジデオキシシチジンが2′,7′−ジクロロフ ルオレセインの5炭素原子に結合し、2′,3′−ジデオキシ−7−デアザグア ノシンおよび2′,3′−ジデオキシ−7−デアザイノシンから成る群から選ば れる前記ジデオキシグアノシンが1′,2′,7′,8′−ジベンゾ−4,7− ジクロロフルオレセインの前記5炭素原子に結合し、そして前記2′,3′−ジ デオキシウリジンがフルオレセインの前記6炭素原子に結合している請求項10 記載の方法。
- 12.前記鋳型核酸を用意し、前記オリゴヌクレオチドプライマーを用意し、ハ イブリッドを形成し、そして伸長する前記工程が、4種の分離した反応混合物の 各々において前記鋳型核酸のためのオリゴヌクレオチドプライマーを用意し、D NAフラグメント個体群の4種のネストシリーズを形成するようにオリゴヌクレ オチドプライマーを別々に伸長する請求項1の記載の方法。
- 13.前記DNAフラグメント個体群を放射活性により標識化し、前記分離工程 がゲル電気泳動によって前記4種のDNAフラグメント個体群の各々を分離して 各バンドが同じ大きさのDNAフラグメントに対応するようにDNAフラグメン ト個体群のバンドの4種の別々の組を形成する請求項12記載の方法。
- 14.前記オリゴヌクレオチドプライマーの各々が標識をもつ請求項12記載の 方法。
- 15.前記オリゴヌクレオチドプライマーの各々のための前記標識が異なる請求 項14記載の方法。
- 16.前記標識がスペクトルにより分割できる1組の蛍光染料から選択される請 求項15記載の方法。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US07/590,218 US5187085A (en) | 1990-09-28 | 1990-09-28 | Nucleic acid sequence analysis with nucleoside-5'-o-(1-thiotriphosphates) |
US590,218 | 1990-09-28 | ||
PCT/US1991/007345 WO1992006219A1 (en) | 1990-09-28 | 1991-09-27 | Improved nucleic acid sequence analysis with nucleoside-5'-0-(1-thiotriphosphates) |
Publications (2)
Publication Number | Publication Date |
---|---|
JPH05507000A true JPH05507000A (ja) | 1993-10-14 |
JPH0661280B2 JPH0661280B2 (ja) | 1994-08-17 |
Family
ID=24361335
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP3517034A Expired - Lifetime JPH0661280B2 (ja) | 1990-09-28 | 1991-09-27 | ヌクレオシド−5’−o−(1−チオトリホスフェート)を用いる改良された核酸シーケンス分析 |
Country Status (6)
Country | Link |
---|---|
US (1) | US5187085A (ja) |
EP (1) | EP0550646B1 (ja) |
JP (1) | JPH0661280B2 (ja) |
AT (1) | ATE191511T1 (ja) |
DE (1) | DE69132097T2 (ja) |
WO (1) | WO1992006219A1 (ja) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2004043819A (ja) * | 1996-05-03 | 2004-02-12 | Applera Corp | 硬質リンカ―を有するエネルギ―転移色素 |
JP2012197269A (ja) * | 2011-03-04 | 2012-10-18 | Utsunomiya Univ | レゾルシノール誘導体 |
Families Citing this family (50)
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US6004446A (en) * | 1984-03-29 | 1999-12-21 | Li-Cor, Inc. | DNA Sequencing |
US4729947A (en) * | 1984-03-29 | 1988-03-08 | The Board Of Regents Of The University Of Nebraska | DNA sequencing |
US6207421B1 (en) | 1984-03-29 | 2001-03-27 | Li-Cor, Inc. | DNA sequencing and DNA terminators |
US6086737A (en) * | 1984-03-29 | 2000-07-11 | Li-Cor, Inc. | Sequencing near infrared and infrared fluorescence labeled DNA for detecting using laser diodes and suitable labels therefor |
US5571388A (en) * | 1984-03-29 | 1996-11-05 | Li-Cor, Inc. | Sequencing near infrared and infrared fluorescense labeled DNA for detecting using laser diodes and suitable labels thereof |
US5360523A (en) * | 1984-03-29 | 1994-11-01 | Li-Cor, Inc. | DNA sequencing |
US5863403A (en) * | 1984-03-29 | 1999-01-26 | The Board Of Regents Of The University Of Nebraska | Digital DNA typing |
EP0660843A1 (en) * | 1992-09-18 | 1995-07-05 | Amoco Corporation | Green fluorescent labeled nucleotides for use in probes |
DE69516325T2 (de) * | 1994-02-22 | 2001-01-18 | Mitsubishi Chem Corp | Oligonukleotid und Verfahren zur Analyse der Basensequenz der Nukleinsäure |
DE4438918A1 (de) * | 1994-11-04 | 1996-05-09 | Hoechst Ag | Modifizierte Oligonukleotide, deren Herstellung sowie deren Verwendung |
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GB9620209D0 (en) * | 1996-09-27 | 1996-11-13 | Cemu Bioteknik Ab | Method of sequencing DNA |
US6017702A (en) * | 1996-12-05 | 2000-01-25 | The Perkin-Elmer Corporation | Chain-termination type nucleic acid sequencing method including 2'-deoxyuridine-5'-triphosphate |
GB9626815D0 (en) | 1996-12-23 | 1997-02-12 | Cemu Bioteknik Ab | Method of sequencing DNA |
AU8162598A (en) | 1997-06-25 | 1999-01-04 | Life Technologies, Inc. | Improved method for isolating and recovering target dna or rna molecules having a desired nucleotide sequence |
AU9476598A (en) * | 1997-09-11 | 1999-03-29 | Seq, Ltd. | Method to make fluorescent nucleotide photoproducts for dna sequencing and analysis |
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US6787305B1 (en) * | 1998-03-13 | 2004-09-07 | Invitrogen Corporation | Compositions and methods for enhanced synthesis of nucleic acid molecules |
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DE3807975C2 (de) * | 1988-03-10 | 2002-03-07 | Karl Otto Greulich | Verfahren zur optischen Charakterisierung von Nukleinsäuren und Oligonukleotiden |
-
1990
- 1990-09-28 US US07/590,218 patent/US5187085A/en not_active Expired - Lifetime
-
1991
- 1991-09-27 AT AT91918268T patent/ATE191511T1/de not_active IP Right Cessation
- 1991-09-27 WO PCT/US1991/007345 patent/WO1992006219A1/en active IP Right Grant
- 1991-09-27 DE DE69132097T patent/DE69132097T2/de not_active Expired - Lifetime
- 1991-09-27 EP EP91918268A patent/EP0550646B1/en not_active Expired - Lifetime
- 1991-09-27 JP JP3517034A patent/JPH0661280B2/ja not_active Expired - Lifetime
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2004043819A (ja) * | 1996-05-03 | 2004-02-12 | Applera Corp | 硬質リンカ―を有するエネルギ―転移色素 |
JP2012197269A (ja) * | 2011-03-04 | 2012-10-18 | Utsunomiya Univ | レゾルシノール誘導体 |
Also Published As
Publication number | Publication date |
---|---|
DE69132097T2 (de) | 2000-10-12 |
JPH0661280B2 (ja) | 1994-08-17 |
EP0550646A4 (en) | 1994-05-25 |
WO1992006219A1 (en) | 1992-04-16 |
DE69132097D1 (de) | 2000-05-11 |
EP0550646A1 (en) | 1993-07-14 |
ATE191511T1 (de) | 2000-04-15 |
EP0550646B1 (en) | 2000-04-05 |
US5187085A (en) | 1993-02-16 |
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