title: GSM2428716: DMSO rep2; Sus scrofa; RNA-Seq
library_construction_protocol: Total RNA was isolated from three untreated (DMSO) and three GW0742-treated primary cartilage cells using Trizol reagent (Invitrogen). The total RNA samples are first treated with DNase I to degrade any possible DNA contamination. Then the mRNA is enriched by using the oligo(dT) magnetic beads. Mixed with the fragmentation buffer, the mRNA is fragmented into short fragments(about 200 bp). Then the first strand of cDNA is synthesized by using random hexamer-primed reverse transcription. Buffer, dNTPs, RNase H and DNA polymerase I are added to synthesize the second strand. The double strand cDNA is purified with magnetic beads. End reparation is then performed. After the previous step, adaptors are ligated to the end of these fragments. Next, ligation products were selected by size and purified on TAE-agarose gel. Finally, the fragments are enriched by PCR amplification, then purified by magnetic beads and dissolved in the appropriate amount of Epstein-Barr solution. During the QC step, Agilent 2100 Bioanaylzer are used to qualify and quantify of the sample library. The library products are ready for sequencing via Ion Proton platform.
attributes.tag: GEO Accession
attributes.value: GSM2428716