GSM2053500: A24_3miRNA; Oryza sativa Indica Group; miRNA-Seq
To construct nine small RNA libraries, three independent samples from each of the three different time points were harvested and individually subjected to small RNA extraction using an Illumina TruSeq Small RNA Preparation kit (Illumina, San Diego, USA). Total RNA was purified through electrophoretic separation with a 15% TBE-Urea denaturing polyacrylamide gel, and small RNA areas (15-30 nt bands) were excised and recovered. The purified small RNAs were hereafter ligated with 5' adaptors, and the ligation products (40-60 nt in length) were purified on a Novex 15% TBE-Urea gel to abandon un-ligated adaptors. Thereafter, a 3' adaptor was ligated to the 5' ligation products. Following isolation and ligation, the small RNAs were reverse transcribed to cDNAs. The purified cDNAs from the nine RNA libraries were sequenced with an Illumina Genome Analyzer II (LC Sciences, Hangzhou, China) according to the manufacturer's protocols. For degradome libraries, polyA-enriched RNA was isolated and ligated to oligonucleotide adaptors containing an MmeI recognition site. After reverse transcription, the ligated sequence was used to generate first-strand cDNA. A short PCR was then used to amplify the cDNA. After digestion, ligation and purification, the final cDNA library was sequenced on an Illumina Genome Analyzer II (LC Sciences, Hangzhou, China) following manufacturer's instruction.
GEO Accession
GSM2053500