GSM4785333: Alex-Mount-Sinai-PB2OE-3; Homo sapiens; RNA-Seq
Total RNA from WT, PB2 KO and PB2 OE hESCs was isolated using a RNeasy Micro kit (Qiagen), following the recommended instructions for on-column DNase digestion. RNA quality and concentrations were measured using an Agilent Bioanalyzer (Agilent RNA 6000 Pico), and samples with RNA integrity number scores higher than 7 were used. Poly(A) mRNA were isolated using NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB E7490S) and cDNA libraries for Illumina nextgeneration sequencing were prepared with NEBNext Ultra Directional RNA Library Prep Kit (NEBE7760S). Three independent samples for each condition were barcoded using NEBNext Index primers 7 (NEB, E7335S), pooled together as one sample, and run on an Illumina platform HiSeq2500 with onesided 75 base reads, with about 30 million reads per sample.