alias: GSM3680184
title: GSM3680184: OCI-Ly1-CONTROL-sg1-01; Homo sapiens; RNA-Seq
library_construction_protocol: RNA extraction according the manufacterer's protocol using the RNAeasy mini Kit (Qiagen) Total RNA was quantified using the Quant-iT™ RiboGreen® RNA Assay Kit and normalized to 5 ng/μl. Following plating, 2 μL of ERCC controls (using a 1:1000 dilution) were spiked into each sample. An aliquot of 200ng for each sample was transferred into library preparation which uses 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, followed by heat fragmentation and cDNA synthesis from the RNA template. The resultant 400bp cDNA then goes through dual-indexed library preparation: 'A' base addition, adapter ligation using P7 adapters, and PCR enrichment using P5 adapters. After enrichment, the libraries were quantified using Quant-iT PicoGreen (1:200 dilution). After normalizing samples to 5 ng/μL, the set was pooled and quantified using the KAPA Library Quantification Kit for Illumina Sequencing Platforms.  The entire process was in a 96-well format and all pipetting is done by either Agilent Bravo or Hamilton Starlet.
identifiers.namespace: GEO
identifiers.id: GSM3680184