公开(公告)号：US11898200B2

公开(公告)日：2024-02-13

申请号：US17014322

申请日：2020-09-08

Applicant: Huaqiao University

Inventor： Philipp Kapranov ,  Huifen Cao ,  Lorena Salazar-García ,  Fan Gao ,  Dongyang Xu ,  Ye Cai ,  Xueer Han ,  Fang Wang ,  Lu Tang

IPC: C12Q1/686 ,  C12Q1/6827

CPC classification number: C12Q1/686 ,  C12Q1/6827

Abstract: The present disclosure discloses a method for detecting single strand breaks (SSBs) in DNA based on the following steps. First, DNA of interest is fragmented with a method that generates 3′ ends that cannot be tailed. Second, the available 3′ ends of the fragmented DNA corresponding to the pre-existing breaks are tailed. Third, SSBs are captured and their positions are identified genome-wide based on the following steps: (1) the tailed fragments are linearly amplified using a chimeric 5′-DNA-RNA-3′ primer; (2) the products of primer extension are tailed at the 3′ ends; (3) the desired products are amplified by PCR with oligonucleotides containing Illumina® adaptor sequences complementary to both tails and subjected to next-generation sequencing (NGS); 4) finally, positions of SSBs are revealed through the analysis of sequencing results.