title: GSM1145199: Endoderm G1/S phase (day6-3 h); Homo sapiens; RNA-Seq
library_construction_protocol: Sequencing libraries were generated by first isolating total RNA using RNeasy Plus Mini Kit (Qiagen) and then purifying mRNA using Dynabeads® Oligo (dT)25 (Invitrogen) according to the manufacturer's instructions. 3' RNA-Seq libraries were prepared as previously described (Beck et al., PLOS One 2010). Briefly, mRNA was heat sheared for 7 minutes to produce an average fragment size range of 300-500 bp, then used to generate cDNA libraries using a custom oligo dT primer containing Illumina-compatible adapter sequence. cDNA fragments were end-repaired and ligated to standard Illumina adapters. Products were PCR amplified for 15 cycles, and purified using Qiagen MinElute columns. Library quality was assessed using Bioanalyzer and Qubit, and sequenced on the Genome Analyzer IIx.