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1. 发明申请

WO2008045575A2 SEQUENCING METHOD 审中-公开
标题翻译：测序方法
公开(公告)号：WO2008045575A2
公开(公告)日：2008-04-17
申请号：PCT/US2007021981
申请日：2007-10-15
申请人： CRAIG J VENTER INST INC , LEVY SAMUEL , GOLDBERG SUSANNE , BEESON KAREN
发明人： LEVY SAMUEL , GOLDBERG SUSANNE , BEESON KAREN
IPC分类号： C12N15/09
CPC分类号： C12Q1/6869 , C12Q2521/301 , C12Q2525/191
摘要： The present invention relates, e.g., to a method for isolating a DNA molecule of interest in a form suitable for sequencing at least a portion of the DNA by a high throughput sequencing method, comprising (a) digesting a double-stranded (ds) DNA molecule with two different restriction enzymes, A and B, to generate a ds form of the DNA molecule of interest, which is bounded by the two restriction enzyme cleavage products, and (b) attaching to each end of the DNA molecule of interest an adaptor molecule which comprises at one end a restriction enzyme cleavage site that is compatible with the restriction enzyme A or the r estriction enzyme B cleavage product, and which also comprises a sequence and/or element that allows the DNA of interest to be sequenced with a high throughput sequencing apparatus. The method can be adapted for sequencing DNA with a variety of high throughput sequencing apparatuses, including machines manufactured by the 454, Illumina (Solexa Sequencing technology) and ABI (SOLiD TM Sequencing technology) companies. A method is also described for sequencing regulatory elements within a cell, comprising subjecting a collection of ds DNA molecules that are enriched for regulatory elements and that are generated by digestion with two restriction enzymes, A and B, which generate sticky ends, to an isolation method of the invention, and sequencing the collection of ds DNA molecules with a high throughput sequencing apparatus.
摘要翻译：本发明涉及例如以适合于通过高通量测序方法测序至少一部分DNA的形式分离感兴趣的DNA分子的方法，其包括（a）消化双链（ds）DNA 分子与两种不同的限制性内切酶A和B连接以产生由两种限制酶切割产物结合的感兴趣的DNA分子的ds形式，和（b）在感兴趣的DNA分子的每个末端连接衔接子所述分子在一个末端包含与限制性酶A或限制性酶B裂解产物相容的限制性酶切割位点，并且还包含允许所关注的DNA以高的顺序测序的序列和/或元件吞吐量测序装置。该方法可适用于用各种高通量测序仪器对DNA进行测序，包括由454，Illumina（Solexa测序技术）和ABI（SOLiD TM测序技术）公司制造的机器。还描述了用于对细胞内的调节元件进行测序的方法，所述方法包括使富集调节元件并且通过用产生粘性末端的两种限制酶A和B消化产生的dsDNA分子的集合进行分离本发明的方法，并用高通量测序仪对ds DNA分子的集合进行测序。

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