Transposase-assisted tagmentation of RNA/DNA hybrid duplexes

eLife

Liting Dong¹  Meiling Zhang¹  Chenxu Zhu²  Xiaoyu Li²  Danyang Yi²  Chengqi Yi²  Bo Lu² 

[1] Peking-Tsinghua Center for Life Sciences, Peking University, Beijing, China;State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing, China;

关键词: RNA/DNA hybrids;    Tn5;    tagmentation;    transposition;    RNA-seq;    epitranscriptome;   

DOI  :  10.7554/eLife.54919

来源: DOAJ

【 摘 要 】

Tn5-mediated transposition of double-strand DNA has been widely utilized in various high-throughput sequencing applications. Here, we report that the Tn5 transposase is also capable of direct tagmentation of RNA/DNA hybrids in vitro. As a proof-of-concept application, we utilized this activity to replace the traditional library construction procedure of RNA sequencing, which contains many laborious and time-consuming processes. Results of Transposase-assisted RNA/DNA hybrids Co-tagmEntation (termed ‘TRACE-seq’) are compared to traditional RNA-seq methods in terms of detected gene number, gene body coverage, gene expression measurement, library complexity, and differential expression analysis. At the meantime, TRACE-seq enables a cost-effective one-tube library construction protocol and hence is more rapid (within 6 hr) and convenient. We expect this tagmentation activity on RNA/DNA hybrids to have broad potentials on RNA biology and chromatin research.

【 授权许可】

Unknown