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SRX3554647: TS4_At_rep1
1 ILLUMINA (Illumina MiSeq) run: 2.6M spots, 129.6M bases, 64.8Mb downloads

Design: kit Illumina Truseq + PEG + HD adapters
Submitted by: CNRS-I2BC
Study: Systematic comparison of small RNA library preparation protocols for next-generation sequencing
show Abstracthide Abstract
Next-generation sequencing technologies have revolutionized the study of small RNAs (sRNAs) on a genome-wide scale. However, classical sRNA library preparation methods introduce serious bias, mainly during adapter ligation steps. Several types of sRNA including plant microRNAs (miRNA), piwi-interacting RNAs (piRNA) in insects, nematodes and mammals, and small interfering RNAs (siRNA) in insects and plants contain a 2'-O-methyl (2'-OMe) modification at their 3' terminal nucleotide. This inhibits 3' adapter ligation and makes library preparation particularly challenging. To reduce bias, the NEBNext kit (New England Biolabs) uses polyethylene glycol (PEG), the NEXTflex V2 kit (BIOO Scientific) uses both randomised adapters and PEG, and the novel SMARTer (Clontech) and CATS (Diagenode) kits avoid ligation altogether. Here we compared these methods with Illumina's classical TruSeq protocol regarding the detection of normal and 2' OMe RNAs. In addition, we modified the TruSeq and NEXTflex protocols to identify conditions that improve performance.
SAMN08354757 • SRS2828007 • All experiments • All runs
Name: TS4_At_rep1
Instrument: Illumina MiSeq
Strategy: miRNA-Seq
Selection: RT-PCR
Layout: SINGLE
Runs: 1 run, 2.6M spots, 129.6M bases, 64.8Mb
Run# of Spots# of BasesSizePublished


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