GSM8995055: Ad-NEXN+high-Pi, rep1; Homo sapiens; RNA-Seq
RNA was extracted and purified using TRIzol Reagent (Invitrogen, 15596026). Its quantity and purity were measured with a NanoDrop ND - 1000. RNA integrity was ensured with a RIN score above 7.0, assessed by Bioanalyzer 2100 and agarose gel electrophoresis. Poly(A) RNA was selected from total RNA using Dynabeads Oligo (dT) 25 - 61005. The selected poly(A) RNA fragments were reverse - transcribed into cDNA by SuperScript™ II Reverse Transcriptase (Invitrogen, cat. 1896649). Then, it was converted into U - labeled second - stranded DNAs. An A - base was added to the ends for adapter ligation. Indexed adapters were ligated to the A - tailed DNA, and products were size - selected with AMPureXP beads. After UDG enzyme (NEB, cat.m0280) treatment, the products were amplified by PCR. The final cDNA library had an average insert size of 300±50 bp.