公开(公告)号：US20140329712A1

公开(公告)日：2014-11-06

申请号：US14347690

申请日：2012-09-28

Applicant: STC.UNM

Inventor： Jeremy Edwards ,  Payman Zarkesh-Ha ,  Steven R.J. Brueck

IPC: C12Q1/68

CPC classification number: C12Q1/6869 ,  C12Q1/6806 ,  C12Q1/6874 ,  G01N27/414 ,  G01N27/4145 ,  Y10T436/143333 ,  C12Q2531/125 ,  C12Q2535/122

Abstract: This disclosure describes, in one aspect, a method for preparing DNA molecule for sequencing. Generally, the method includes fragmenting the DNA molecule into double-stranded fragments; amplifying at least a portion of the double-stranded fragments; circularizing the fragments so that the first end of the fragment comprises a first loop connecting the strands and the second end of the fragment comprises a second loop connecting the strands; annealing a first sequencing primer to the first loop oriented to sequence at least a portion of one strand of the fragment; and annealing a second sequencing primer to the second loop oriented to sequence at least a portion of the other strand of the fragment. In another aspect, this disclosure describes a method for sequencing a DNA molecule. Generally, the method includes fragmenting the DNA molecule into double-stranded fragments; amplifying at least a portion of the double-stranded fragments; circularizing the fragments so that the first end of the fragment comprises a first loop connecting the strands and the second end of the fragment comprises a second loop connecting the strands; and sequencing at least one of the DNA strands.

Abstract translation: 本公开一方面描述了用于测序的DNA分子的制备方法。 通常，该方法包括将DNA分子片段化成双链片段; 扩增至少一部分双链片段; 使片段环化，使得片段的第一端包含连接线束的第一环和片段的第二端包括连接线的第二环; 将第一测序引物退火至所述第一环，其定向为序列所述片段的一条链的至少一部分; 以及将第二测序引物退火至所述第二环，其定向为序列所述片段的另一条链的至少一部分。 在另一方面，本公开描述了用于测序DNA分子的方法。 通常，该方法包括将DNA分子片段化成双链片段; 扩增至少一部分双链片段; 使片段环化，使得片段的第一端包含连接线束的第一环和片段的第二端包括连接线的第二环; 并测序至少一个DNA链。