title: RNA-seq of human GM12878 and JURKAT cells treated with aphidicolin and Sendai virus against untreated controls
design: RNA-seq of human GM12878 and JURKAT cells treated with aphidicolin and Sendai virus against untreated controls
library_construction_protocol: At the end of 24 hours, cells were washed with cold 1x PBS twice. Cells were collected in RLT buffer (Qiagen) supplemented with dTT as suggested by RNeasy mini RNA extraction protocol. GM12878 and JURKAT cells were cultured with RPMI media supplemented with 10% FBS 37C with 5% CO2. When cells are 80% confluent, 1 uM of aphidicolin was added to the media and incubated for 12 hours at 37C. Sendai Virus or media was added to the media and incubated for and additional of 12 hours at 37C. RNA was isolated using RNeasy mini kit (Qiagen) according to manufacturer's instructions. DNA was removed by 2h treatment with TURBO DNase (Thermo Fisher Scientific) and RNA was cleaned up using RNA Clean & Concentrator-25 kit according to the supplier's instructions (Zymo Research). Library prep was performed according to TruSeq® Stranded mRNA kit (Illumina) protocol.