Transposase assisted tagmentation of RNA/DNA hybrid duplexes
- Peking University, China
Abstract
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 6h) and convenient. We expect this tagmentation activity on RNA/DNA hybrids to have broad potentials on RNA biology and chromatin research.
Data availability
High-throughput sequence data has been deposited in GEO under accession code GSE143422. REVIEW: To review GEO accession GSE143422, please go to https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE143422 and enter token cfilsuwgplodnap into the box.
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Transposase assisted tagmentation of RNA/DNA hybrid duplexesNCBI Gene Expression Omnibus, GSE143422.
Article and author information
Author details
Funding
National Natural Science Foundation of China (31861143026)
- Chengqi Yi
National Natural Science Foundation of China (91740112)
- Chengqi Yi
Ministry of Science and Technology of the People's Republic of China (2019YFA0110900)
- Chengqi Yi
Ministry of Science and Technology of the People's Republic of China (2019YFA0802201)
- Chengqi Yi
National Natural Science Foundation of China (21825701)
- Chengqi Yi
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Copyright
© 2020, Lu et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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Transposase assisted tagmentation of RNA/DNA hybrid duplexes
eLife 9:e54919.
https://doi.org/10.7554/eLife.54919
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