title: GSM8212671: WT kidney, hypoxia, 12 h,rep1; Mus musculus; RNA-Seq
library_construction_protocol: Total RNA was extracted using the CTAB method. 1 μg of total RNA was used for library construction After being purified from total RNA by using poly-T oligo-attached magnetic beads, the mRNA was cut into fragments using divalent cations under elevated temperature in First Strand Synthesis Reaction Buffer(5X). Random hexamer primer and M-MuLV Reverse Transcriptase were used to synthesize the first strand cDNA. PCR was performed to synthesize the second strand cDNA. Then the remaining overhangs were converted into blunt ends. After adenylation of 3' ends of DNA fragments, Adaptor with hairpin loop structure was ligated to prepare for hybridization. AMPure XP system (Beckman Coulter, Beverly, USA) was used to select 370~420 bp cDNA fragments, which were then amplified by PCR and purified by AMPure XP beads again to get the library.