| Autor: |
Giraldez, Maria D, Spengler, Ryan M, Etheridge, Alton, Godoy, Paula M, Barczak, Andrea J, Srinivasan, Srimeenakshi, De Hoff, Peter L, Tanriverdi, Kahraman, Courtright, Amanda, Lu, Shulin, Khoory, Joseph, Rubio, Renee, Baxter, David, Driedonks, Tom A P, Buermans, Henk P J, Nolte-'t Hoen, Esther N M, Jiang, Hui, Wang, Kai, Ghiran, Ionita, Wang, Yaoyu E |
| Zdroj: |
Nature Biotechnology; Aug2022, Vol. 40 Issue 8, p746-757, 12p |
| Abstrakt: |
Systematic evaluation of library preparation methods for small RNA-seq identifies reproducible and accurate methods. RNA-seq is increasingly used for quantitative profiling of small RNAs (for example, microRNAs, piRNAs and snoRNAs) in diverse sample types, including isolated cells, tissues and cell-free biofluids. The accuracy and reproducibility of the currently used small RNA-seq library preparation methods have not been systematically tested. Here we report results obtained by a consortium of nine labs that independently sequenced reference, 'ground truth' samples of synthetic small RNAs and human plasma-derived RNA. We assessed three commercially available library preparation methods that use adapters of defined sequence and six methods using adapters with degenerate bases. Both protocol- and sequence-specific biases were identified, including biases that reduced the ability of small RNA-seq to accurately measure adenosine-to-inosine editing in microRNAs. We found that these biases were mitigated by library preparation methods that incorporate adapters with degenerate bases. MicroRNA relative quantification between samples using small RNA-seq was accurate and reproducible across laboratories and methods. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
| Externí odkaz: |
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