JP2001521389A - アダプターに基づく配列分析の改良 - Google Patents
アダプターに基づく配列分析の改良Info
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- JP2001521389A JP2001521389A JP54426098A JP54426098A JP2001521389A JP 2001521389 A JP2001521389 A JP 2001521389A JP 54426098 A JP54426098 A JP 54426098A JP 54426098 A JP54426098 A JP 54426098A JP 2001521389 A JP2001521389 A JP 2001521389A
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- 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
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Abstract
Description
Claims (1)
- 【特許請求の範囲】 1.標的ポリヌクレオチドのヌクレオチド配列を決定するための方法であって 、該方法が、 (a)(i)第1鎖および第2鎖を含む二本鎖標的ポリヌクレオチドであって、ここ で、該第1鎖は、5'-ヒドロキシル基を有する該ポリヌクレオチドの一方の末端 で終結し、そして該第2鎖は、3'-リン酸基を有する同じ該ポリヌクレオチド末 端で終結する、二本鎖標的ポリヌクレオチド、および(ii)第1鎖および第2鎖を 含む二本鎖ポリヌクレオチドアダプターであって、ここで、該第1鎖は、3'-ヒ ドロキシルブロッキング基を有する該アダプターの一方の末端で終結し、そして 該第2鎖は、5'-リン酸基を有する同じ該アダプター末端で終結する、二本鎖ポ リヌクレオチドアダプター、を提供する工程; (b)該標的ポリヌクレオチドの該第2鎖が、該アダプターの該第2鎖にライゲ ートされるように、該標的ポリヌクレオチド末端を該アダプター末端にライゲー トし、1本鎖のライゲートした標的-アダプター付加物を形成する、工程; (c)該付加物における該3'-ヒドロキシルブロッキング基を3'-ヒドロキシル基 に変換し、そして該付加物における該5'-ヒドロキシル基を5'-リン酸基に変換す る工程であって、該変換が、同時にまたはいずれかの順で連続して行われる、工 程; (d)該標的ポリヌクレオチドの該第2鎖を、工程(c)後の該アダプターの該第2 鎖にライゲートし、2本鎖のライゲートした標的-アダプター付加物を形成する 、工程;および (e)工程(d)に続いて、該標的ポリヌクレオチドにおける1つ以上のヌクレオチ ドを同定する工程、 を包含する、方法。 2.請求項1に記載の方法であって、前記アダプターが、該アダプターの前記 末端に対する配向を有するII型エンドヌクレアーゼ認識部位を有し、そのため、 工程(d)の前記2本鎖のライゲートした標的-アダプター付加物が該エンドヌクレ アーゼと反応する場合、(i)該エンドヌクレアーゼの切断部位が、該アダプター の該末端を越えて少なくとも1つのヌクレオチドを越えたところで生じ、そして (ii)該エンドヌクレアーゼの該切断のパターンが、該アダプターの該末端に相補 的な末端を有する短縮型標的ポリヌクレオチドフラグメントを生成するために効 果的であり、そして該方法が、該認識部位に特異的なII型エンドヌクレアーゼで 該2本鎖のライゲートした付加物を切断し、1つ以上のヌクレオチドによって短 縮された短縮型標的ポリヌクレオチドフラグメントを産生する工程をさらに包含 する、方法。 3.請求項2に記載の方法であって、ここで前記切断工程後、前記短縮型標的 ポリヌクレオチドフラグメントが、該切断工程によって生じる5'-末端リン酸基 を除去するために処理される、方法。 4.請求項1〜3のいずれか1項に記載の方法であって、ここで前記3'-ブロ ッキング基がリン酸基であり、そして該3'-ブロッキング基を3'-ヒドロキシル基 に変換する前記工程が、ホスファターゼを使用することによって達成される、方 法。 5.請求項1〜3のいずれか1項に記載の方法であって、ここで前記3'-ブロ ッキング基を3'-ヒドロキシル基に変換する前記工程が、前記アダプターの前記 第2鎖を、(i)置換された鎖の配列と同じ配列を有し、そして(ii)3'-ブロッキン グ基よりも3'-ヒドロキシル基を含む、新しい第2鎖と置換する工程を包含する 、方法。 6.請求項1〜5のいずれか1項に記載の方法であって、前記アダプターが、 以下の式を有する、方法: 5'-p(N)n(N)q-3' z(N')r-5' または p(N)q-3' 3'-z(N)n(N')r-5' ここで、NおよびN'は相補的ヌクレオチドであり、pはリン酸基であり、zは3'ブ ロッキング基であり、nは2から5までの全部の整数であり、qは8以上の整数で あり、そしてrは8以上の整数でありそしてqと同じまたは異なり得る。 7.請求項6に記載の方法であって、ここでzがリン酸基であり、qが12〜50の 範囲であり、そしてrが12〜50の範囲である、方法。 8.請求項1〜7のいずれか1項に記載の方法であって、前記標的ポリヌクレ オチドおよび前記ポリヌクレオチドアダプターの前記末端が、互いに相補的であ る5'-オーバーハングを含む、方法。 9.請求項1〜7のいずれか1項に記載の方法であって、前記標的ポリヌクレ オチドおよび前記ポリヌクレオチドアダプターの前記末端が、互いに相補的であ る3'-オーバーハングを含む、方法。 10.前記ポリヌクレオチドが、固相支持体に付着される、請求項1〜9のい ずれか1項に記載の方法。 11.工程(b)〜(e)が1回以上繰り返される、請求項1〜10のいずれか1項 に記載の方法。 12.請求項1〜11のいずれか1項に記載の方法における使用のための組成 物であって、該組成物が、以下の式の二本鎖核酸アダプターを含む、組成物: 5'-p(N)n(N)q-3' z(N')r-5' または p(N)q-3' 3'-z(N)n(N')r-5' ここで、NおよびN'は相補的ヌクレオチドであり、pはリン酸基であり、zは3'ブ ロッキング基であり、nは2から5までの全部の整数であり、qは8以上の整数で あり、そしてrは8以上の整数でありそしてqと同じまたは異なり得る。 13.請求項12に記載の組成物であって、ここでzがリン酸基であり、qが12 〜50の範囲であり、そしてrが12〜50の範囲である、組成物。 14.請求項12または13に記載の組成物であって、ここで前記標的ポリヌ クレオチドおよび前記ポリヌクレオチドアダプターの前記末端が、互いに相補的 である5'-オーバーハングを含む、組成物。 15.請求項12または13に記載の組成物であって、ここで前記標的ポリヌ クレオチドおよび前記ポリヌクレオチドアダプターの前記末端が、互いに相補的 である3'-オーバーハングを含む、組成物。 16.実質的に同一の二本鎖ポリヌクレオチドの均一な集団を付着しているマ イクロパーティクルであって、各ポリヌクレオチドが、同じ末端によって該マイ クロパーティクルに付着し、そして各ポリヌクレオチドが、該マイクロパーティ クルと離れた方の末端で脱リン酸化された5'-または3'-突出鎖を有する、マイク ロパーティクル。 17.前記突出鎖が2〜5ヌクレオチド長である、請求項16に記載のマイク ロパーティクル。 18.前記実質的に同一のポリヌクレオチドが、実質的に同一のcDNAである、 請求項16または請求項17に記載のマイクロパーティクル。 19.前記マイクロパーティクルが、CPGマイクロパーティクル、GMAマイクロ パーティクル、またはポリスチレンマイクロパーティクルである、請求項16〜 18のいずれか1項に記載のマイクロパーティクル。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
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US08/842,608 US5888737A (en) | 1997-04-15 | 1997-04-15 | Adaptor-based sequence analysis |
US08/842,608 | 1997-04-15 | ||
PCT/US1998/007592 WO1998046621A1 (en) | 1997-04-15 | 1998-04-14 | Improvements in adaptor-based sequence analysis |
Publications (2)
Publication Number | Publication Date |
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JP2001521389A true JP2001521389A (ja) | 2001-11-06 |
JP4110216B2 JP4110216B2 (ja) | 2008-07-02 |
Family
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Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP54426098A Expired - Lifetime JP4110216B2 (ja) | 1997-04-15 | 1998-04-14 | アダプターに基づく配列分析の改良 |
Country Status (7)
Country | Link |
---|---|
US (2) | US5888737A (ja) |
EP (1) | EP0975655B1 (ja) |
JP (1) | JP4110216B2 (ja) |
AU (1) | AU737331B2 (ja) |
CA (1) | CA2286400A1 (ja) |
DE (1) | DE69836809T2 (ja) |
WO (1) | WO1998046621A1 (ja) |
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US5093245A (en) * | 1988-01-26 | 1992-03-03 | Applied Biosystems | Labeling by simultaneous ligation and restriction |
US5237016A (en) * | 1989-01-05 | 1993-08-17 | Siska Diagnostics, Inc. | End-attachment of oligonucleotides to polyacrylamide solid supports for capture and detection of nucleic acids |
US5270185A (en) * | 1989-04-21 | 1993-12-14 | Hoffmann-La Roche Inc. | High-efficiency cloning of CDNA |
US5516663A (en) * | 1990-01-26 | 1996-05-14 | Abbott Laboratories | Ligase chain reaction with endonuclease IV correction and contamination control |
CA2036946C (en) * | 1990-04-06 | 2001-10-16 | Kenneth V. Deugau | Indexing linkers |
WO1992020697A1 (en) * | 1991-05-10 | 1992-11-26 | Hybridon, Inc. | 3'-end blocked oligonucleotides |
WO1993005183A1 (en) * | 1991-09-09 | 1993-03-18 | Baylor College Of Medicine | Method and device for rapid dna or rna sequencing determination by a base addition sequencing scheme |
GB9214873D0 (en) * | 1992-07-13 | 1992-08-26 | Medical Res Council | Process for categorising nucleotide sequence populations |
WO1994002634A1 (en) * | 1992-07-24 | 1994-02-03 | University Of South Australia | Amplification and detection process |
US5503980A (en) * | 1992-11-06 | 1996-04-02 | Trustees Of Boston University | Positional sequencing by hybridization |
GB9401200D0 (en) * | 1994-01-21 | 1994-03-16 | Medical Res Council | Sequencing of nucleic acids |
US5552278A (en) * | 1994-04-04 | 1996-09-03 | Spectragen, Inc. | DNA sequencing by stepwise ligation and cleavage |
CZ397998A3 (cs) * | 1996-06-06 | 1999-07-14 | Lynx Therapeutics, Inc. | Způsob sekvenční analýzy ligací kódovaného adaptoru |
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- 1998-04-14 AU AU71213/98A patent/AU737331B2/en not_active Ceased
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- 1998-04-14 JP JP54426098A patent/JP4110216B2/ja not_active Expired - Lifetime
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EP0975655A1 (en) | 2000-02-02 |
DE69836809T2 (de) | 2007-10-11 |
JP4110216B2 (ja) | 2008-07-02 |
CA2286400A1 (en) | 1998-10-22 |
WO1998046621A1 (en) | 1998-10-22 |
US5888737A (en) | 1999-03-30 |
EP0975655B1 (en) | 2007-01-03 |
US6175002B1 (en) | 2001-01-16 |
AU737331B2 (en) | 2001-08-16 |
AU7121398A (en) | 1998-11-11 |
EP0975655A4 (en) | 2004-03-31 |
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