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Updated: Jan 18, 2026
Improving Small RNA-seq: Less Bias and Better Detection of 2'-O-Methyl RNAs
Published on: September 16, 2019
Improving Small RNA-seq: Less Bias and Better Detection of 2'-O-Methyl RNAs
Erwin L van Dijk1, Evangelia Eleftheriou2, Claude Thermes2
1Institute for Integrative Biology of the Cell, UMR9198, CNRS CEA Univ Paris-Sud, Université Paris-Saclay; erwin.vandijk@i2bc.paris-saclay.fr.
Abstract:
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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