title: GSM2856388: KD-SHRNA-18-3_S12_L002; Homo sapiens; RNA-Seq
library_construction_protocol: RNA samples from KD siRNA, SCR SiRNA (72 hours post-transfection), KD shRNA and SCR shRNA cells (in triplicates) were prepared using the RNeasy Mini kit (Qiagen). Libraries were generated using the TruSeq Stranded mRNA Library Prep Kit (Illumina). RNA was quantified using Qubit® RNA BR Assay Kit and if necessary diluted to 25-100 ng in 12.5 ul.  This was followed by RNA quality check to establish the RIN (RNA Integrity Number) using the Agilent RNA ScreenTape system. RNA was then processed on the automated Illumina Neoprep system following the Library prep TruSeq Stranded mRNA Library Prep Kit for NeoPrep. Libraries were normalized to 10 nM by the Neoprep. Each library quantity was checked again using Qubit DNA HS Assay Kit and 2 ul of each library were pooled together into a single tube.  This pool of 16 samples was checked on the Agilent High Sensitivity D1000 ScreenTape to ensure the libraries were the correct size at 300 bp. The pooled sample was diluted to 4 nM.