GSM2053622: HG_METFORMIN_REP2; Homo sapiens; RNA-Seq
After the incubation, total RNA was isolated using TRIZOL. Genomic DNA contamination was removed by DNase treatment (Qiagen). mRNA-Seq libraries were prepared according to Illumina mRNA-Seq library preparation kit (cat # RS-100-0801). Briefly, 8 µg of total RNA was taken for mRNA isolation using poly-T oligo-attached magnetic beads. Poly-A mRNA was fragmented with the presence of divalent cations under elevated temperatures. The mRNA fragments were reverse transcribed via SuperScript II (Invitrogen) with random primers. The second strand was synthesized with addition of RNaseH and DNA Pol I. Double stranded complementary DNA (dscDNAs) were end-repaired, followed by 3’ adenylation and ligation to universal adapters. The ligated cDNA library was separated on a 2.0% agarose gel. Library cDNA fragments in the range of 200 bp ± 10 bp were isolated by gel purification (Qiagen) and amplified with 15 cycles of PCR
GEO Accession
GSM2053622