title: GSM1860500: LR3_miRNA; Chiloscyllium plagiosum; miRNA-Seq
library_construction_protocol: The total RNA in the Chiloscyllium plagiosum liver was extracted using the Total RNA Purification Kit (LC Sciences) following the manufacturer’s protocol, and the deep sequencing of small RNAs extracted from Chiloscyllium plagiosum liver was performed at LC Sciences using Illumina’s Genome analyzer II platform. Small RNAs were size-fractionated to isolate RNA fragments 15–50 nt long and ligated to SRA5’ adapter , and 41–76 nt long RNA fragments were isolated. This step was followed by SRA3’ adapter ligation and The 64–99 nt long RNA fragments were isolated. The RNAs were converted reverse transcribed to single-stranded cDNAs and then PCR amplified for 20 cycles using Illumina’s small RNA primer set, and cDNA in the 80-115 bp range that contained miRNA was isolated. This fraction was eluted, precipitated and quantified on Nanodrop (Thermo Scientific). The concentration of cDNA sample was adjusted to 10nM and 10 µl was used for sequencing reaction.. Purified cDNA was used for cluster generation on Illumina’s Cluster Station and sequenced using Illumina GAIIx platform Raw sequencing reads were obtained using the Illumina Pipeline v1.5 software following sequencing image analysis by the Pipeline Firecrest Module and base-calling by the Pipeline Bustard Module.
attributes.tag: GEO Accession
attributes.value: GSM1860500