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Article Dans Une Revue Journal of visualized experiments : JoVE Année : 2019

Improving Small RNA-seq: Less Bias and Better Detection of 2'-O-Methyl RNAs

Résumé

The study of small RNAs (sRNAs) by next-generation sequencing (NGS) is challenged by bias issues during library preparation. Several types of sRNA such as plant microRNAs (miRNAs) carry a 2'-O-methyl (2'-OMe) modification at their 3' terminal nucleotide. This modification adds another difficulty as it inhibits 3' adapter ligation. We previously demonstrated that modified versions of the 'TruSeq (TS)' protocol have less bias and an improved detection of 2'-OMe RNAs. Here we describe in detail protocol 'TS5', which showed the best overall performance. TS5 can be followed either using homemade reagents or reagents from the TS kit, with equal performance.
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Dates et versions

hal-02307637 , version 1 (07-10-2019)

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Erwin L. van Dijk, Evangelia Eleftheriou, Claude Thermes. Improving Small RNA-seq: Less Bias and Better Detection of 2'-O-Methyl RNAs. Journal of visualized experiments : JoVE, 2019, 151, ⟨10.3791/60056⟩. ⟨hal-02307637⟩
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