title: GSM8760960: Homo sapiens, human pluripotent stem cells, control blank, Day3; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA was extracted using the EasyPure® RNA Kit, quality was assessed using the 5300 Bioanalyzer (Agilent) and quantified with the ND-2000 (NanoDrop Technologies). The mRNA fragmentation was performed using divalent cations and high temperature. Priming was performed using Random Primers. First strand cDNA and the second-strand cDNA were synthesized. The purified double-stranded cDNA was then treated to repair both ends and add a dA-tailing in one reaction, followed by a T-A ligation to add adaptors to both ends. Size selection of Adaptor-ligated DNA was then performed using DNA Clean Beads. Each sample was then amplified by PCR using P5 and P7 primers and the PCR products were validated. Then libraries with different indexs were multiplexed and loaded on an Illumina instrument for sequencing using a 2x150 paired-end (PE) configuration according to manufacturer's instructions.