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    Single molecule sequencing with two distinct chemistry steps
    5.
    发明授权
    Single molecule sequencing with two distinct chemistry steps 有权
    具有两个不同化学步骤的单分子测序

    公开(公告)号：US09587275B2

    公开(公告)日：2017-03-07

    申请号：US14848136

    申请日：2015-09-08

    申请人： Pacific Biosciences of California, Inc.

    发明人： Robin Emig ,  Lei Jia ,  Jeremiah Hanes ,  Lubomir Sebo

    IPC分类号： C12Q1/68 ,  C07H19/10 ,  C07H19/11 ,  C07H19/20 ,  C07H19/213

    CPC分类号： C12Q1/6869 ,  C07H19/10 ,  C07H19/11 ,  C07H19/20 ,  C07H19/213 ,  C12Q1/6874 ,  C12Q2521/301 ,  C12Q2525/186 ,  C12Q2563/107 ,  C12Q2537/157 ,  C12Q2521/319 ,  C12Q2521/525

    摘要： Methods, Compositions, and Systems are provided for nucleic acid sequencing where the sequential incorporation of nucleotides uses two distinct chemical steps. A plurality of nucleotide analogs, each having a labeled leaving group at its 3′ hydroxyl can be sequentially added to a growing strand in the presence of a selective cleaving activity that cleaves the 3′ hydroxyl leaving group preferentially after it has been incorporated. The selective cleaving agent can comprise an exonuclease activity, and the exonuclease activity can be a polymerase-associated exonuclease activity. Nucleotide analogs having labels on both a cleavable polyphosphate portion and on a 3′ hydroxyl leaving group can provide signals characteristic of nucleotide analog incorporation. Systems having illumination optics, collection optics, and substrates observe signals from the labels as they are being incorporated into a growing nucleic acid strand, allowing for the sequencing of template nucleic acids.

    摘要翻译： 提供了用于核酸测序的方法，组合物和系统，其中连续掺入核苷酸使用两个不同的化学步骤。 在选择性切割活性的存在下，可以将多个在其3'羟基上具有标记的离去基团的核苷酸类似物依次加入到生长链中，该选择性裂解活性优选在其引入后优先切割3'羟基离去基团。 选择性切割剂可以包含核酸外切酶活性，并且核酸外切酶活性可以是聚合酶相关的外切核酸酶活性。 在可切割聚磷酸酯部分和3'羟基离去基团上具有标记的核苷酸类似物可以提供核苷酸类似物掺入特征的信号。 具有照明光学器件，收集光学元件和底物的系统在标记被并入生长的核酸链中观察信号，允许对模板核酸进行测序。