阅读说明：本技术 测序文库的构建方法、测序方法及试剂盒和应用 (Construction method of sequencing library, sequencing method, kit and application ) 是由 周荣芳 黄标 刘晨 周晏秋 黄金 田志坚 于 2019-09-20 设计创作，主要内容包括：提供了一种测序文库的构建方法、测序方法及试剂盒和应用。该测序文库的构建方法包括将线性双链DNA样本进行损伤修复和末端修复,以便获得经修复的DNA样本；将所述经修复的DNA样本进行同源重组,以便获得环状双链DNA分子；以环状双链DNA分子变性的环状单链DNA分子为模板,进行滚环扩增,以便获得RCA扩增产物；RCA扩增产物与测序接头连接,以便获得所述测序文库。还提供了测序方法、试剂盒、制备环状双链DNA分子的方法以及同源重组在RCA扩增中的应用。(Provides a construction method, a sequencing method, a kit and application of a sequencing library. The construction method of the sequencing library comprises the steps of carrying out damage repair and end repair on a linear double-stranded DNA sample so as to obtain a repaired DNA sample; subjecting the repaired DNA sample to homologous recombination to obtain circular double-stranded DNA molecules; performing rolling circle amplification by using the circular single-stranded DNA molecule denatured by the circular double-stranded DNA molecule as a template so as to obtain an RCA amplification product; the RCA amplification products are ligated to sequencing adaptors in order to obtain the sequencing library. Also provided are sequencing methods, kits, methods of preparing circular double stranded DNA molecules and the use of homologous recombination in RCA amplification.)