Transposase-assisted tagmentation of RNA/DNA hybrid duplexes.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 32701057.
- Also identified by DOI 10.7554/eLife.54919 and PMC identifier 7402673.
- Licence recorded as CC BY.
- The licence permits redistribution, so the abstract is shown in full and the full text is available from the publisher.
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 <u>T</u>ransposase-assisted <u>R</u>NA/DN<u>A</u> hybrids <u>C</u>o-tagm<u>E</u>ntation (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.
Medical subject headings
- DNA
- RNA
- Sequence Analysis
- Transposases