title: GSM4307129: MDA-MB-231, Inducible GFP Overexpression + Doxorubicin treatment for 48h replicate 1; Homo sapiens; RNA-Seq
library_construction_protocol: RNA-seq libraries were generated from two biological replicates for each condition.  The total RNA was extracted using the RNAeasy kit.  The extracted RNA with an RNA integrity number (RIN) of >7.0 was used for mRNA purification. The mRNA was purified using oligo-dT beads (TruSeq RNA Sample Preparation Kit, Illumina) from 1 ug of intact total RNA. The purified mRNA was fragmented at 90°C in the presence of divalent cations. The fragments were reverse transcribed using random hexamers and Superscript II Reverse Transcriptase (Life Technologies). The sequencing library was prepared by 5' and 3' adapter ligation, after end-repair and the addition of an 'A' base and SPRI cleanup. The prepared cDNA library was amplified using PCR for the enrichment of the adapter-ligated fragments. The individual libraries were quantified using a Qubit fluorometer and validated for quality with a Bioanalyzer (Agilent Technologies). SPRI-based (Beckman Coulter Agencourt Ampure XP)-based purification was used to clean after each enzymatic reaction.