GSM2441047: hES-N2G2 day 12 control; Homo sapiens; RNA-Seq
Total RNA was extracted with Trizol The Illumina TruSeq RNA Library Prep Kit v2 (Illumina) was used following manufacturer’s instructions. The mRNA was then fragmented and copied into first strand cDNA using reverse transcriptase and random primers. The 3' end of the cDNA was then adenylated and adapters were ligated. One of the ligated adapters had a 6-nt barcode that was unique for each sample that allowed us to sequence more than one sample in each lane of HiSeq Flow Cell (Illumina). The products were purified and enriched by PCR to create the final cDNA library. Final libraries were checked for quality and quantity by TapeStation (Agilent Technologies) and qPCR using Kapa’s Library Quantification kit for Illumina sequencing platforms (Kapa Biosystems). The libraries were clustered on the cBot (Illumina) and sequenced 4 samples per lane on a 50 cycle single end on a HiSeq 2500 (Illumina) in High Output mode using version 4 reagents according to manufacturers recommended protocols.
GEO Accession
GSM2441047