alias: GSM2890014
title: GSM2890014: H929; Homo sapiens; RNA-Seq
library_construction_protocol: Cells from cell lines were harvested following confirmation of a cell viability above 95% using the NucleoCounter NC-250 instrument (Chemometec, Alleroed, Denmark). RNA was isolated using the Allprep DNA/RNA/miRNA universal kit (Qiagen, Hilden, Germany) according to the manufacturer's instructions. on-column DNase treatment was performed using 10 µL lyophilized DNase I (1500 kU) in 70 µL buffer RDD and incubated at 15 min. at room temperature. One µg of total RNA was rRNA depleted using the Ribo-Zero rRNA Removal Kit (Human, Mouse, Rat) (Epicentre, Madison, WI, USA) according to the manufacturer's instructions followed by a purification step using AMPure XP Beads (Beckman Coulter, Brea, CA, USA). Sequencing libraries were generated using the ScriptSeq v2 RNA-Seq Library Preparation Kit (Epicentre) according to the manufacturer's instructions with 12 PCR cycles for amplification. Purification was performed using AMPure XP Beads (Beckman Coulter). The final library was quality controlled on the 2100 Bioanalyzer (Agilent Technologies) and quantified using the KAPA library quantification kit (Kapa Biosystems, Wilmington, MA, USA) according to the manufacturer's protocol. High-throughput sequencing was performed on the HiSeq 4000 system (Illumina, San Diego, CA, USA) at the Beijing Genomics Institute (BGI) using the 100 paired-end sequencing protocol. Nine samples were pooled for each lane on the HiSeq 4000.
identifiers.namespace: GEO
identifiers.id: GSM2890014