GSM9105642: sgNTC-3; Homo sapiens; RNA-Seq
Total RNA was extracted using standard TRIzol-based methods (or as per sample-specific protocol). RNA quality was assessed via Agilent Bioanalyzer and quantified with Qubit 2.0. mRNA was purified from total RNA using poly-T oligo-attached magnetic beads. Fragmentation was performed in First Strand Synthesis Reaction Buffer under elevated temperature. First strand cDNA was synthesized with random hexamer primers and M-MuLV reverse transcriptase. Second strand synthesis was performed using DNA Polymerase I and RNase H. After end repair and A-tailing, adapter ligation was done, followed by PCR amplification and size selection (~370–420 bp) using AMPure XP beads.