title: GSM3764690: SHI1_1; Homo sapiens; RNA-Seq
library_construction_protocol: Trizol RNA preparation Total RNA was quantified using the Quant-iT RiboGreen RNA Assay Kit and normalized to 5 ng/µl. 200 ng of each sample was transferred into library preparation which was an automated variant of the Illumina TruSeq Stranded mRNA Sample Preparation Kit. This method preserves strand orientation of the RNA transcript. It uses oligo dT beads to select mRNA from the total RNA sample. It is followed by heat fragmentation and cDNA synthesis from the RNA template. The resultant cDNA then goes through library preparation (end repair, base 'A' addition, adapter ligation, and enrichment) using indexed adapters for multiplexing. After enrichment the libraries were quantified with qPCR using the KAPA Library Quantification Kit for Illumina Sequencing Platforms and then pooled equimolarly. The entire process was in 96-well format and all pipetting was done by either Agilent Bravo or Hamilton Starlet. Pooled libraries were normalized to 2 nM and denatured using 0.1 N NaOH prior to sequencing.