JP7137595B2 - 化学組成物とそれを利用する方法 - Google Patents
化学組成物とそれを利用する方法 Download PDFInfo
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- JP7137595B2 JP7137595B2 JP2020115005A JP2020115005A JP7137595B2 JP 7137595 B2 JP7137595 B2 JP 7137595B2 JP 2020115005 A JP2020115005 A JP 2020115005A JP 2020115005 A JP2020115005 A JP 2020115005A JP 7137595 B2 JP7137595 B2 JP 7137595B2
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- 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
- C12Q2563/00—Nucleic acid detection characterized by the use of physical, structural and functional properties
- C12Q2563/179—Nucleic acid detection characterized by the use of physical, structural and functional properties the label being a nucleic acid
-
- 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
- C12Q2565/00—Nucleic acid analysis characterised by mode or means of detection
- C12Q2565/10—Detection mode being characterised by the assay principle
- C12Q2565/101—Interaction between at least two labels
-
- 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
- C12Q2565/00—Nucleic acid analysis characterised by mode or means of detection
- C12Q2565/50—Detection characterised by immobilisation to a surface
- C12Q2565/518—Detection characterised by immobilisation to a surface characterised by the immobilisation of the nucleic acid sample or target
Description
本出願は、2016年11月21日に出願されたアメリカ合衆国仮出願第62/424,887号と、2017年2月10日に出願されたアメリカ合衆国仮出願第62/457,237号と、2017年7月24日に出願されたアメリカ合衆国仮出願第62/536,147号の優先権と恩恵を主張する。上記特許出願のそれぞれの内容は、前記全体が参照によって本明細書に組み込まれている。
本出願は、EFS-Webを通じてASCII形式で提出された配列リストを含有しており、前記全体が参照によって本明細書に組み込まれている。このASCIIのコピーは2017年11月20日に作成されてNATE033-001WO_SeqList_ST25.txtと名づけられており、サイズは19,351バイトである。
Claims (25)
- 少なくとも6個の二次核酸分子にハイブリダイズした一次核酸分子を含むプローブであって、
前記一次核酸分子が、第1のドメインおよび第2のドメインを含み、前記少なくとも6個の二次核酸分子が、前記一次核酸分子の前記第2のドメインにハイブリダイズし、
前記一次核酸分子が、前記第1のドメインと前記第2のドメインとの間に少なくとも1個の光切断可能なリンカーを含み、
前記二次核酸分子のそれぞれは、少なくとも5個の三次核酸分子にハイブリダイズし、
各二次核酸分子が、第1のドメインおよび第2のドメインを含み、前記第1のドメインが前記一次核酸分子にハイブリダイズし、前記第2のドメインが少なくとも5個の三次核酸分子にハイブリダイズし、
前記二次核酸分子のそれぞれが、その第1のドメインと第2のドメインとの間に位置する少なくとも1個の光切断可能なリンカーを含み、
前記プローブが少なくとも30個の検出可能な標識を含むように、前記三次核酸分子のそれぞれは、少なくとも1個の検出可能な標識を含む、プローブ。 - 前記第1のドメインが、14個のヌクレオチドを含む、請求項1に記載のプローブ。
- 前記一次核酸分子が、100~80個のヌクレオチドを含む、請求項2に記載のプローブ。
- 各二次核酸分子が、80~90個のヌクレオチドを含む、請求項1に記載のプローブ。
- 各三次核酸分子が、10~20個のヌクレオチドを含む、請求項1に記載のプローブ。
- 各三次核酸分子が、15個のヌクレオチドを含む、請求項5に記載のプローブ。
- 前記少なくとも30個の検出可能な標識のすべてが、同じ発光スペクトルを有する、請求項1に記載のプローブ。
- 前記30個の検出可能な標識の少なくとも1個が、第1の発光スペクトルを有し、前記30個の検出可能な標識の少なくとも1個が、第2の発光スペクトルを有する、請求項1に記載のプローブ。
- 前記第1の発光スペクトルと前記第2の発光スペクトルが、スペクトル分解可能である、請求項8に記載のプローブ。
- 前記少なくとも1個の検出可能な標識が、蛍光標識である、請求項1に記載のプローブ。
- サンプル中の標的核酸をシークエンシングする方法であって、
(a)プローブの標的結合ドメインが標的核酸と結合するような条件下で、サンプルを標的結合ドメインとバーコードドメインを含むシークエンシング用プローブと接触させること;
(b)プローブが前記シークエンシング用プローブのバーコードドメインに結合するような条件下で、サンプルを請求項1のプローブと接触させること;
(c)前記プローブの検出可能な標識を検出し、それによりサンプル中の標的核酸をシークエンシングすること、
を含む、方法。 - サンプル中の標的核酸をシークエンシングする方法であって、
(a)プローブの標的結合ドメインが標的核酸と結合するような条件下で、サンプルを標的結合ドメインとバーコードドメインを含むシークエンシング用プローブと接触させること;
(b)プローブが前記シークエンシング用プローブのバーコードドメインに結合するような条件下で、サンプルを請求項1の第1プローブと接触させること;
(c)前記第1プローブの検出可能な標識を検出すること;
(d)前記第1プローブの検出可能な標識を除去すること;
(e)プローブが前記シークエンシング用プローブのバーコードドメインに結合するような条件下で、サンプルを請求項1に記載の少なくとも第2のプローブと接触させること;
(f)前記第2のプローブの検出可能な標識を検出し、それによりサンプル中の標的核酸をシークエンシングすること、
を含む、方法。 - 複数の二次核酸分子にハイブリダイズした一次核酸分子を含むプローブであって、
前記一次核酸分子が、第1のドメインおよび第2のドメインを含み、前記第2のドメインが、複数の二次核酸分子にハイブリダイズし、
前記一次核酸分子が、前記第1のドメインと前記第2のドメインとの間に少なくとも1個の光切断可能なリンカーを含み、
前記二次核酸分子のそれぞれが、複数の三次核酸分子にハイブリダイズし、
各二次核酸分子が、第1のドメインおよび第2のドメインを含み、前記第1のドメインが一次核酸分子にハイブリダイズし、そして、前記第2のドメインが複数の三次核酸分子にハイブリダイズし、
前記二次核酸分子のそれぞれが、その第1のドメインと第2のドメインとの間に位置する少なくとも1個の光切断可能なリンカーを含み、
前記複数のうちの少なくとも1個の三次核酸分子が、少なくとも5個の検出可能な標識を含み、そして、
前記プローブが少なくとも36個の検出可能な標識を含むように、前記複数のうちの残りの三次核酸分子のそれぞれが、少なくとも1個の検出可能な標識を含む、プローブ。 - 前記一次核酸分子が、少なくとも4個の二次核酸分子にハイブリダイズする、請求項13に記載のプローブ。
- 前記二次核酸分子のそれぞれが、少なくとも5個の三次核酸分子にハイブリダイズする、請求項13に記載のプローブ。
- 前記三次核酸分子の少なくとも1個が標識されたオリゴヌクレオチドである、請求項13に記載のプローブ。
- 少なくとも4個の三次核酸分子が、標識されたオリゴヌクレオチドである、請求項13に記載のプローブ。
- 少なくとも5個の検出可能な標識が、5個の標識されたオリゴヌクレオチドを介して少なくとも1個の三次核酸分子にハイブリダイズする、請求項13に記載のプローブ。
- 少なくとも5個の三次核酸分子が、標識されたオリゴヌクレオチドである4個の三次核酸分子、および、少なくとも5個の検出可能な標識を含む1個の三次核酸分子を含み、ここで、前記少なくとも5個の検出可能な標識が、5個の標識されたオリゴヌクレオチドを介して1個の三次核酸分子にハイブリダイズする、請求項15に記載のプローブ。
- 前記少なくとも1個の三次核酸分子の少なくとも5個の検出可能な標識が、残りの三次核酸分子の少なくとも1個の検出可能な標識とは異なる発光スペクトルを有する、請求項13に記載のプローブ。
- 複数の二次核酸分子のうちの少なくとも1個が、複数の三次核酸分子にハイブリダイズし、ここで、複数のうちの少なくとも1個の三次核酸分子が、第1の発光スペクトルを有する少なくとも5個の検出可能な標識を含み、複数の残りの三次核酸分子のそれぞれは、第2の発光スペクトルを有する少なくとも1個の検出可能な標識を含み、そして
複数の二次核酸分子のうちの少なくとも1個が、複数の三次核酸分子にハイブリダイズし、複数のうちの少なくとも1個の三次核酸分子が、第2の発光スペクトルを有する少なくとも5個の検出可能な標識を含み、複数のうちの残りの三次核酸分子のそれぞれが、第1の発光スペクトルを有する少なくとも1個の検出可能な標識を含む、請求項13に記載のプローブ。 - 前記一次核酸分子が、少なくとも1個の修飾されたヌクレオチドまたはヌクレオチド類似体を含む、請求項13に記載のプローブ。
- 少なくとも1つの検出可能な標識が、蛍光標識である、請求項13に記載のプローブ。
- サンプル中の標的核酸をシークエンシングする方法であって、
(a)プローブの標的結合ドメインが標的核酸と結合するような条件下で、サンプルを標的結合ドメインとバーコードドメインを含むシークエンシング用プローブと接触させること;
(b)プローブが前記シークエンシング用プローブのバーコードドメインに結合するような条件下で、サンプルを請求項13に記載のプローブと接触させること;
(c)プローブの検出可能な標識を検出し、それによりサンプル中の標的核酸をシークエンシングすること、
を含む、方法。 - サンプル中の標的核酸をシークエンシングする方法であって、
(a)プローブの標的結合ドメインが標的核酸と結合するような条件下で、サンプルを標的結合ドメインとバーコードドメインを含むシークエンシング用プローブと接触させること;
(b)プローブが前記シークエンシング用プローブのバーコードドメインに結合するような条件下で、サンプルを請求項13の第1のプローブと接触させること;
(c)第1のプローブの検出可能な標識を検出すること;
(d)第1のプローブの検出可能な標識を除去すること;
(e)プローブが前記シークエンシング用プローブのバーコードドメインに結合するような条件下で、サンプルを請求項13に記載の少なくとも第2のプローブと接触させること;
(f)第2のプローブの検出可能な標識を検出し、それによりサンプル中の標的核酸をシークエンシングすること、
を含む、方法。
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