GSM2373499: BCBL-1_DMSO; Homo sapiens; RNA-Seq
Total RNA was isolated using the TRIzol Reagent (Invitrogen) from MCF-7 parental and TMR cells treated with or without 4-hydroxytamoxifen for 24 hours. Indexed RNA-Seq libraries were prepared from 200 ng total RNA using the KAPA Stranded mRNA-Seq Kit (Kapa Biosystems, Inc.) according to the manufacturer’s protocol. Poly-adenylated mRNA was purified by binding to oligo(dT) beads and to remove rRNA, which was followed by fragmentation by incubation at 94ºC in the presence of magnesium. Double-stranded cDNA was then generated by random-primed first-strand synthesis and second strand synthesis in the presence of dUTP for strand marking. Subsequently, the cDNA was 3’-A tailed and indexed, Illumina-compatible adapters were ligated. The libraries were then enriched by high-fidelity PCR amplification (13 cycles) with KAPA HiFi HotStart DNA Polymerase and adapter-specific primers.
GEO Accession
GSM2373499