阅读说明：本技术 用于检测低频突变的靶向富集方法和试剂盒 (Targeted enrichment method and kit for detecting low-frequency mutation ) 是由 杨林 高雅 蒲丹丹 张海萍 程云阳 陈芳 蒋慧 于 2017-07-21 设计创作，主要内容包括：本发明提供了一种用于检测低频突变的靶向富集方法和试剂盒。该方法包括：在末端转移酶的作用下,在单链DNA和/或双链DNA每条链的3’端加上一段单一碱基的单聚核苷酸尾巴；以上游特异性引物和第一通用引物对突变目标位点所在的区域进行第一次PCR扩增,其中第一通用引物包括5’端的测序引物序列1和3’端的与单聚核苷酸尾巴互补的连续单一碱基；以下游特异性引物和第二通用引物对第一次PCR扩增的产物进行第二次PCR扩增,其中第二通用引物包括第一通用引物的测序引物序列1。(The invention provides a targeted enrichment method and a kit for detecting low-frequency mutation. The method comprises the following steps: under the action of terminal transferase, adding a single-nucleotide tail with single base at the 3' end of each strand of the single-stranded DNA and/or the double-stranded DNA; performing first PCR amplification on a region where a mutation target site is located by using an upstream specific primer and a first universal primer, wherein the first universal primer comprises a sequencing primer sequence 1 at a 5 'end and continuous single bases which are complementary with a single nucleotide tail at a 3' end; performing a second PCR amplification on the product of the first PCR amplification with a downstream specific primer and a second universal primer, wherein the second universal primer comprises the sequencing primer sequence 1 of the first universal primer.)