GSM3381918: E2; Homo sapiens; RNA-Seq
Cells were washed with PBS and lysed in QIAzol Lysis Reagent (Qiagen) by vortexing and incubating for 5 min at 20°C. Lysates were stored at -80°C until RNA extraction. RNA was extracted with the miRNeasy Micro Kit (Qiagen) according to the manufacturer's instructions. RNA concentration was determined on a Nanodrop 2000 spectrophotometer (Thermo Fisher Scientific) and RNA quality was controlled on a 2100 Bioanalyzer instrument (Agilent Technologies) using an Agilent RNA 6000 Pico Kit. RNA integrity numbers (RIN) were 8.5±0.3 (mean±SD). Libraries were prepared in one batch with the TruSeq Stranded mRNA HT kit (Illumina) with Dual Indexing adapters and Ambion ERCC Spike-In Control (Thermo Fisher Scientific). Samples were allocated to sequencing indexes and -lanes in a balanced fashion to control for potential batch effects (7-8 samples/lane). Library concentration was determined on a Qubit 2.0 Fluorometer (Thermo Fisher Scientific). Library size and quality were measured on a 2100 Bioanalyzer instrument (Agilent Technologies) using an Agilent High Sensitivity DNA Kit. Libraries were quantified with the KAPA Library Quantification Kit (KAPA Biosystems). Sequencing was performed on a HiSeq 2500 Sequencing Platform (Illumina; High Output Run) with 76 nt paired-end reads.
GEO
GSM3381918
GEO Accession
GSM3381918