公开(公告)号：WO2010027484A2

公开(公告)日：2010-03-11

申请号：PCT/US2009004993

申请日：2009-09-04

Applicant: PACIFIC BIOSCIENCES CALIFORNIA ,  PATEL PRANAV ,  KORLACH JONAS ,  BIBILLO ARKADIUSZ ,  BJORNSON KEITH ,  HANES JEREMIAH

Inventor： PATEL PRANAV ,  KORLACH JONAS ,  BIBILLO ARKADIUSZ ,  BJORNSON KEITH ,  HANES JEREMIAH

IPC: C12Q1/68 ,  C12N15/11 ,  C12Q1/25

CPC classification number: C12Q1/6869 ,  C12N9/1252 ,  C12Q2527/113 ,  C12Q2521/101 ,  C12Q2527/137 ,  C12Q2537/161 ,  C12Q2525/101 ,  C12Q2563/107

Abstract: Provided are methods for enhanced sequencing of nucleic acid templates. Also provided are reaction conditions that increase branching fractions during polymerization reactions. Also provided are compositions comprising modified recombinant polymerases that exhibit branching fractions that are higher than the branching fractions of the polymerases from which they were derived. Provided are compositions comprising modified recombinant polymerases that exhibit delayed translocation relative to the polymerases from which they were derived. Also provided are compositions comprising modified recombinant polymerases that exhibit increased nucleotide or nucleotide analog residence time at an active site of the polymerase. Provided are methods for generating polymerases with the aforementioned phenotypes and methods of using such polymerases to sequence a DNA template or make a DNA. Also provided are methods and nucleic acid sequencing systems for determining which labeled nucleotide is incorporated at a site during a template-dependent polymerization reaction.

Abstract translation: 提供了用于增强核酸模板测序的方法。 还提供了在聚合反应期间增加支化馏分的反应条件。 还提供了包含改性重组聚合酶的组合物，其表现出高于其衍生的聚合酶的支化部分的支化部分。 提供了包含修饰的重组聚合酶的组合物，其相对于衍生自其的聚合酶表现出延迟的移位。 还提供了包含修饰的重组聚合酶的组合物，其在聚合酶的活性位点表现出增加的核苷酸或核苷酸类似物停留时间。 提供了使用上述表型产生聚合酶的方法和使用这种聚合酶来顺序DNA模板或制备DNA的方法。 还提供了用于在模板依赖性聚合反应期间确定哪个标记的核苷酸被掺入位点的方法和核酸测序系统。

公开(公告)号：WO2010027484A3

公开(公告)日：2010-06-24

申请号：PCT/US2009004993

申请日：2009-09-04

Applicant: PACIFIC BIOSCIENCES CALIFORNIA ,  PATEL PRANAV ,  KORLACH JONAS ,  BIBILLO ARKADIUSZ ,  BJORNSON KEITH ,  HANES JEREMIAH

Inventor： PATEL PRANAV ,  KORLACH JONAS ,  BIBILLO ARKADIUSZ ,  BJORNSON KEITH ,  HANES JEREMIAH

IPC: C12Q1/68 ,  C12N15/11 ,  C12Q1/25

CPC classification number: C12Q1/6869 ,  C12N9/1252 ,  C12Q2527/113 ,  C12Q2521/101 ,  C12Q2527/137 ,  C12Q2537/161 ,  C12Q2525/101 ,  C12Q2563/107

Abstract: Provided are methods for enhanced sequencing of nucleic acid templates. Also provided are reaction conditions that increase branching fractions during polymerization reactions. Also provided are compositions comprising modified recombinant polymerases that exhibit branching fractions that are higher than the branching fractions of the polymerases from which they were derived. Provided are compositions comprising modified recombinant polymerases that exhibit delayed translocation relative to the polymerases from which they were derived. Also provided are compositions comprising modified recombinant polymerases that exhibit increased nucleotide or nucleotide analog residence time at an active site of the polymerase. Provided are methods for generating polymerases with the aforementioned phenotypes and methods of using such polymerases to sequence a DNA template or make a DNA. Also provided are methods and nucleic acid sequencing systems for determining which labeled nucleotide is incorporated at a site during a template-dependent polymerization reaction.

Abstract translation: 提供了用于增强核酸模板测序的方法。 还提供了在聚合反应期间增加支化部分的反应条件。 还提供了包含修饰的重组聚合酶的组合物，其表现出比衍生它们的聚合酶的支化部分更高的支化部分。 提供了包含相对于衍生它们的聚合酶显示延迟易位的修饰的重组聚合酶的组合物。 还提供了包含修饰的重组聚合酶的组合物，所述修饰的重组聚合酶在聚合酶的活性位点显示增加的核苷酸或核苷酸类似物停留时间 提供了产生具有上述表型的聚合酶的方法和使用这些聚合酶测序DNA模板或制备DNA的方法。 还提供了用于确定哪些标记的核苷酸在模板依赖性聚合反应期间在位点处掺入的方法和核酸测序系统。

公开(公告)号：WO2009120374A3

公开(公告)日：2009-12-17

申请号：PCT/US2009001930

申请日：2009-03-27

Applicant: PACIFIC BIOSCIENCES CALIFORNIA ,  PATEL PRANAV ,  BJORNSON KEITH ,  TRAVERS KEVIN ,  HEINER CHERYL

Inventor： PATEL PRANAV ,  BJORNSON KEITH ,  TRAVERS KEVIN ,  HEINER CHERYL

IPC: C12N15/10 ,  C07H21/00 ,  C12N15/11 ,  C12Q1/68

CPC classification number: C12N15/10 ,  C12N15/65 ,  C12N15/66

Abstract: Provided are methods and compositions for the production of linear single- stranded nucleic acids, which can be used as templates in high-throughput sequencing systems. Also provided are methods and compositions for the production of closed single- stranded nucleic acid loops, which can be used as templates in high-throughput sequencing systems.

Abstract translation: 提供用于生产线性单链核酸的方法和组合物，其可以在高通量测序系统中用作模板。 还提供了用于生产封闭的单链核酸环的方法和组合物，其可以用作高通量测序系统中的模板。

公开(公告)号：AU2009288696A1

公开(公告)日：2010-03-11

申请号：AU2009288696

申请日：2009-09-04

Applicant: PACIFIC BIOSCIENCES CALIFORNIA INC

Inventor： PATEL PRANAV ,  KORLACH JONAS ,  BIBILLO ARKADIUSZ ,  BJORNSON KEITH ,  HANES JEREMIAH

IPC: C12Q1/68 ,  C12N15/11 ,  C12Q1/25

Abstract: Provided are methods for enhanced sequencing of nucleic acid templates. Also provided are reaction conditions that increase branching fractions during polymerization reactions. Also provided are compositions comprising modified recombinant polymerases that exhibit branching fractions that are higher than the branching fractions of the polymerases from which they were derived. Provided are compositions comprising modified recombinant polymerases that exhibit delayed translocation relative to the polymerases from which they were derived. Also provided are compositions comprising modified recombinant polymerases that exhibit increased nucleotide or nucleotide analog residence time at an active site of the polymerase. Provided are methods for generating polymerases with the aforementioned phenotypes and methods of using such polymerases to sequence a DNA template or make a DNA. Also provided are methods and nucleic acid sequencing systems for determining which labeled nucleotide is incorporated at a site during a template-dependent polymerization reaction.

公开(公告)号：CA2735979A1

公开(公告)日：2010-03-11

申请号：CA2735979

申请日：2009-09-04

Applicant: PACIFIC BIOSCIENCES CALIFORNIA

Inventor： PATEL PRANAV ,  KORLACH JONAS ,  BIBILLO ARKADIUSZ ,  BJORNSON KEITH ,  HANES JEREMIAH

IPC: C12Q1/68 ,  C12N15/11 ,  C12Q1/25

Abstract: Provided are methods for enhanced sequencing of nucleic acid templates. Also provided are reaction conditions that increase branching fractions during polymerization reactions. Also provided are compositions comprising modified recombinant polymerases that exhibit branching fractions that are higher than the branching fractions of the polymerases from which they were derived. Provided are compositions comprising modified recombinant polymerases that exhibit delayed translocation relative to the polymerases from which they were derived. Also provided are compositions comprising modified recombinant polymerases that exhibit increased nucleotide or nucleotide analog residence time at an active site of the polymerase. Provided are methods for generating polymerases with the aforementioned phenotypes and methods of using such polymerases to sequence a DNA template or make a DNA. Also provided are methods and nucleic acid sequencing systems for determining which labeled nucleotide is incorporated at a site during a template-dependent polymerization reaction.