GSM8266831: t7_168h_rep2; Mus musculus; RNA-Seq
cells were trypsinised, MEFs were adsorbed on non-treated plasticware for 20 minutes. Cells were washed once in PBS and lysed in TRIzol. Total RNAs were purified by pheno-chloroform extraction and precipitated. Messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers. Then the second strand cDNA was synthesized using dUTP. The directional library was ready after end repair, A-tailing, adapter ligation, size selection, USER enzyme digestion, amplification, and purification, following NOVOGENE library construction workflow. NovogenestrandedRNAkit:250~300bpinsertstrandedspecificlibrary(directional)