title: GSM2266099: RE1; Capra hircus; ncRNA-Seq
library_construction_protocol: Total RNA was extracted from the six samples using the TRIzol reagent (Invitrogen, CA, USA) following the manufacturer's procedure. The quantity and purity of total RNA were analyzed with Bioanalyzer 2100 (Agilent, CA, USA) and RNA 6000 Nano LabChip Kit (Agilent, CA, USA) with RIN number> 7.0. At the same time, a new library was constructed by mixing all the six total RNAs with equal amount, and it was used to decrease false positive dentification of circRNA. Approximately 10 ug of total RNA representing a specic adipose type was used to deplete ribosomal RNA according to the manuscript of the Epicentre Ribo-Zero Gold Kit (Illumina, San Diego, USA). Following purication, the RNAs were treated with RNase R for 20 min at 37 ℃ (Epicentre Technologies, Madison, WI), and then the poly (A)- or poly(A)+ RNA fractions is fragmented into small pieces using divalent cations under elevated temperature. Then the cleaved RNA fragments were reverse-transcribed to create the final cDNA library in accordance with the protocol for the RNA-Seq sample preparation kit (Illumina, San Diego, USA), the average insert size for the paired-end libraries was 250 bp (±50 bp).  Paired-end sequencing on an Illumina Hiseq2500 (LC Sceiences, USA) following the vendor's recommended protocol.
attributes.tag: GEO Accession
attributes.value: GSM2266099