中国海洋大学学报（自然科学版）2024，Vol.54Issue(9)：157-164,8.DOI:10.16441/j.cnki.hdxb.20230181
重叠延伸PCR生成长重复DNA序列及其在纳米孔测序中的应用
Overlap-Extension PCR Generating Long Repetitive DNA Sequence and Its Application in Nanopore Sequencing
摘要
Abstract
Nanopore sequencing of short DNA fragments suffers from low pore utilization and poor data quality.It is urgent to establish a method of obtaining the sequence of short DNA fragments.In this study,PCR was performed at constant 72 ℃ using 1 nmol/L fully complementary primers and short DNA fragments as the template.The overlapping extension of short DNA fragments ultimately generated long tandem repeats,5～20 times longer than the original.The long DNA constructs can be sequenced directly with the accurate original sequence obtained simply by aligning only a few reads.The developed overlap-extension method improved the one read accuracy of nanopore sequencing to 99.28％,which plays an important role in improving the accuracy of nanopore sequencing.关键词
短链DNA/重复序列/纳米孔测序/重叠延伸/高精度Key words
short DNA/repetitive sequences/nanopore sequencing/overlap-extension/high accuracy分类
生物科学引用本文复制引用
宋子婷,程冰晓,胡坤灵,安然,梁兴国..重叠延伸PCR生成长重复DNA序列及其在纳米孔测序中的应用[J].中国海洋大学学报（自然科学版）,2024,54(9):157-164,8.基金项目
国家自然科学基金项目(32102064) （32102064）
国家重点研究发展计划项目(2019YFD0901701)资助 Supported by the National Natural Science Foundation of China(32102064) （2019YFD0901701）
the National Key Research and Development Program of China(2019YFD0901701) （2019YFD0901701）
