GSM1695913: human ts 1 RT-; Homo sapiens; RNA-Seq
Total RNA was extracted using a miRNeasy Mini kit. Libraries were prepared using the TruSeq Stranded mRNA Sample Prep Kit v2 according to the manufacturer’s protocol.  Poly (A)-positive RNA was purified from 250-500 mg of total RNA using streptavidin-coated magnetic beads (AMPure XP) and subsequently fragmented to ~300 bp. cDNA was synthesized using reverse transcriptase (SuperScript II, Invitrogen) and random primers. The strand-specific RNA-Seq library preparation was based on the incorporation of dUTP in place of dTTP in the second strand of the cDNA. Double-stranded DNA was further used for library preparation. Such dsDNA was subjected to A-tailing and ligation of the barcoded Truseq adapters. Library amplification was performed by PCR on the size selected fragments using the primer cocktail supplied in the kit. Final libraries were analyzed using Agilent DNA 1000 chip to estimate the quantity and check size distribution, and they were quantified by qPCR using the KAPA Library Quantification Kit (KapaBiosystems). Sequencing was done with a Illumina HiSeq 2000 sequencer in a paired-end design according to the manufacturer’s instructions.
GEO Accession
GSM1695913