Transposase-assisted tagmentation of RNA/DNA hybrid duplexes
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 re...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
eLife Sciences Publications Ltd
2020-07-01
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| Series: | eLife |
| Subjects: | |
| Online Access: | https://elifesciences.org/articles/54919 |
| _version_ | 1811180965547999232 |
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| author | Bo Lu Liting Dong Danyang Yi Meiling Zhang Chenxu Zhu Xiaoyu Li Chengqi Yi |
| author_facet | Bo Lu Liting Dong Danyang Yi Meiling Zhang Chenxu Zhu Xiaoyu Li Chengqi Yi |
| author_sort | Bo Lu |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-11T09:12:05Z |
| format | Article |
| id | doaj.art-c16288b554134615b49b9f73a509d518 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-11T09:12:05Z |
| publishDate | 2020-07-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-c16288b554134615b49b9f73a509d5182022-12-22T04:32:29ZengeLife Sciences Publications LtdeLife2050-084X2020-07-01910.7554/eLife.54919Transposase-assisted tagmentation of RNA/DNA hybrid duplexesBo Lu0https://orcid.org/0000-0002-5852-0477Liting Dong1https://orcid.org/0000-0001-8396-374XDanyang Yi2Meiling Zhang3Chenxu Zhu4https://orcid.org/0000-0003-4216-6562Xiaoyu Li5Chengqi Yi6https://orcid.org/0000-0003-2540-9729State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing, China; Peking-Tsinghua Center for Life Sciences, Peking University, Beijing, ChinaState Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing, China; Peking-Tsinghua Center for Life Sciences, Peking University, Beijing, ChinaState Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing, ChinaState Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing, ChinaState Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing, ChinaState Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing, ChinaState Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing, China; Peking-Tsinghua Center for Life Sciences, Peking University, Beijing, China; Department of Chemical Biology and Synthetic and Functional Biomolecules Center, College of Chemistry and Molecular Engineering, Peking University, Beijing, ChinaTn5-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.https://elifesciences.org/articles/54919RNA/DNA hybridsTn5tagmentationtranspositionRNA-seqepitranscriptome |
| spellingShingle | Bo Lu Liting Dong Danyang Yi Meiling Zhang Chenxu Zhu Xiaoyu Li Chengqi Yi Transposase-assisted tagmentation of RNA/DNA hybrid duplexes eLife RNA/DNA hybrids Tn5 tagmentation transposition RNA-seq epitranscriptome |
| title | Transposase-assisted tagmentation of RNA/DNA hybrid duplexes |
| title_full | Transposase-assisted tagmentation of RNA/DNA hybrid duplexes |
| title_fullStr | Transposase-assisted tagmentation of RNA/DNA hybrid duplexes |
| title_full_unstemmed | Transposase-assisted tagmentation of RNA/DNA hybrid duplexes |
| title_short | Transposase-assisted tagmentation of RNA/DNA hybrid duplexes |
| title_sort | transposase assisted tagmentation of rna dna hybrid duplexes |
| topic | RNA/DNA hybrids Tn5 tagmentation transposition RNA-seq epitranscriptome |
| url | https://elifesciences.org/articles/54919 |
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