title: Illumina HiSeq 2500 paired end sequencing; The impact of XAB2 on gene expression and pre-mRNA splicing
design: The impact of XAB2 on gene expression and pre-mRNA splicing
library_construction_protocol: Naive mESCs transcfected with dsRNA targeting XAB2 or negative siRNA control (scrable, AllStars Negative (Qiagen)) at a final concentration 50nM. Naive mESCs were cultured on gelatinized tissue culture dishes in medium containing Dulbecco's modified Eagle's medium (DMEM) supplemented with 15% fetal bovine serum (FBS), 50μg mL-1 streptomycin, 50 U mL-1 penicillin (Gibco), 2mM L-glutamine (Gibco), 1% non-essential amino acids (Gibco), 0.1mM β-Mercaptoethanol (Applichem), 1μM MEK inhibitor PD0325901 (Selleck), 3μM GSK inhibitor CHIR99021 (Selleck). Total RNA was isolated from cells using a Total RNA isolation kit (Qiagen) as described by the manufacturer. Standard TruSeq Stranded mRNA