GSM8567944: Ischemic_0min_2; Rattus norvegicus; RNA-Seq
Traditional phenol/chloroform-based extraction was performed for the RNA extraction. Messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, thefirst strand cDNA was synthesized using random hexamer primers, followed by the second strand cDNA synthesis usingeither dUTP for directional library or dTTP for non-directional library. For the non-directional library, it was ready after end repair, A-tailing, adapter ligation, size selection, amplification,and purification. For the directional library, it was ready after end repair, A-tailing, adapter ligation, size selection, USER enzymedigestion, amplification, and purification. The library was checked with Qubit and real-time PCR for quantification and bioanalyzer for size distributiondetection. Quantified libraries will be pooled and sequenced on Illumina platforms, according to effective libraryconcentration and data amount.