title: GSM8163209: DNFB+OVA4; Mus musculus; RNA-Seq
library_construction_protocol: RNA was harvested using Trizol reagent. Illumina Truseq RNA Sample Prep Kit (Cat#FC-122-1001) was used to total RNA extraction. Total RNA was used as input material for the RNA sample preparations.Briefly, mRNA was purified from total RNA using poly-T oligo-attached magneticbeads. Fragmentation was carried out using divalent cations under elevated temperature inFirst Strand Synthesis Reaction Buffer(5X). First strand cDNA was synthesized usingrandom hexamer primer and M-MuLV Reverse Transcriptase(RNase H-). Second strandcDNA synthesis was subsequently performed using DNA Polymerase I and RNase HRemaining overhangs were converted into blunt ends via exonuclease/polymerase activities.After adenylation of 3'ends of DNA fragments, Adaptor with hairpin loop structure wereligated to prepare for hybridization. In order to select cDNA fragments of preferentially370~420 bp in length, the library fragments were purified with AMPure XP system(Beckman Coulter, Beverly, USA). Then PCR was performed with Phusion High-FidelityDNA polymerase, Universal PCR primers and Index (X) Primer. At last, PCR products werepurified (AMPure XP system) and library quality was assessed on the Agilent Bioanalyzer2100 system.