title: GSM1701812: hESC SOX2/3 KD2; Homo sapiens; RNA-Seq
library_construction_protocol: collecting the cells in Trizol Reagent ( Invitrogen, 15596026) The cDNA libraries were constructed following the TruSeq™ RNA Sample Preparation Guide (Illumina). Briefly, total RNA was isolated with Trizol reagent, and the polyA RNA was isolated using the RNA Purification Beads (Illumina). The mRNAs were fragmented by incubation in Elute, Prime, Fragment Mix at 94℃ for 8min to obtain 120-200bp inserts.  Fist strand cDNA was synthesized with SuperScript II Reverse Transcriptase (Invitrogen) using random primer, and Ampure XP beads are used to isolated the double‐stranded (ds) cDNA synthesized by Second Strand Master Mix. The adapter was ligated to the A‐Tailing  fragment, and 12 cycles of PCR was performed to enrich those DNA fragments that have adapter molecules on both ends and to amplify the amount of DNA in the library.  Purified libraries were quantified by Qubit® 2.0 Fluorometer and  validated by Agilent 2100 bioanalyzer to confirm the insert size and calculate the mole   concentration. Cluster was generated by cBot with the library diluted to 10 pM and then were sequenced on the Illumina Genome Analyzer IIx for 75 cycles. The library construction and sequencing was performed at Shanghai Biotechnology Corporation.
attributes.tag: GEO Accession
attributes.value: GSM1701812