GSM1577739: Effector 3h; Homo sapiens; RNA-Seq
RNA was extracted with Aurum™ Total RNA Mini Kit (Bio-Rad) A modified dUTP method was used for preparation of RNA-Seq libraries[27]. Briefly, mRNA was purified from total RNA by Dynabeads® mRNA Purification Kit (Invitrogen) and reverse-transcribed with SuperScript® III (Invitrogen). The second strand was created by DNA Polymerase 1 (NEB) in the presence of RNase H and a dNTP mix containing dUTP instead dTTP. The cDNA was fragmented on a Covaris sonicator, and end repair and A-tailing were performed using the Quick Blunting kit and exo- Klenow fragment of DNA polymerase I, respectively (NEB). Illumina genomic adapters were ligated, and the sample was size-fractionated (~170–400 bp) on E-Gel EX 2% agarose (Invitrogen). After uracil-dna glycosylase (UDG) treatment (NEB) and a final PCR amplification step with Illumina primers (18 cycles), the cDNA libraries were size-fractionated (~175-400 bp) on a 2% agarose gel, quantified using a Qubit 2.0 fluorometer and dsDNA HS kit (Invitrogen), and sent to sequencing on a Illumina HiSeq 2500 sequencing system
GEO Accession
GSM1577739