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Nanopore Sequencing Accurately Identifies the Mutagenic DNA Lesion O 6 ‐Carboxymethyl Guanine and Reveals Its Behavior in Replication
Author(s) -
Wang Yu,
Patil Kiran M.,
Yan Shuanghong,
Zhang Panke,
Guo Weiming,
Wang Yuqin,
Chen HongYuan,
Gillingham Dennis,
Huang Shuo
Publication year - 2019
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.201902521
Subject(s) - dna , nanopore , dna sequencing , nanopore sequencing , microbiology and biotechnology , guanine , dna replication , chemistry , biophysics , biology , computational biology , genetics , nanotechnology , gene , nucleotide , materials science
O 6 ‐carboxymethylguanine (O 6 ‐CMG) is a highly mutagenic alkylation product of DNA, triggering transition mutations relevant to gastrointestinal cancer. However, precise localization of a single O 6 ‐CMG with conventional sequencing platforms is challenging. Here nanopore sequencing (NPS), which directly senses single DNA bases according to their physiochemical properties, was employed to detect O 6 ‐CMG. A unique O 6 ‐CMG signal was observed during NPS and a single‐event call accuracy of >95 % was achieved. Moreover, O 6 ‐CMG was found to be a replication obstacle for Phi29 DNA polymerase (Phi29 DNAP), suggesting this lesion could cause DNA sequencing biases in next generation sequencing (NGS) approaches.