title: GSM6873827: MDA-MB-231 cells, DMSO, Rep2; Homo sapiens; RNA-Seq
library_construction_protocol: Cell counts were determined using C-Chip disposable hemocytometers (Bulldog bio, DHC-N01) and equalized across all samples before lysis and RNA extraction. Total RNA from 1E6 cells per replicate was isolated using RNeasy96 kit (Qiagen). External RNA Controls Consortium (ERCC) ExFoldRNA Spike-in Control Mixes (Invitrogen #4456740) (4 µL/sample, diluted at 1:100, ERCC User Guide, Table 4) were added after cell lysis step. The extraction was then continued according to manufacturer's instructions and eluted in 50 µL nuclease-free water. Total RNA was quantified using QubitTM RNA Broad Range Assay kit (Invitrogen) and analyzed on Agilent 4200 TapeStation for integrity. Samples with the RNA Integrity Number (RIN) above 9.0 and normalized to 500 ng total RNA for library preparation using TruSeq® stranded mRNA sample prep kit (Illumina, #20020595) following manufacturer's protocols. After library preparation, samples were quantified using QubitTM assay, checked fragment sizes on Agilent 4200 TapeStation, normalized and pooled for sequencing.