GSM3374813: Wt_rep_A; Homo sapiens; RNA-Seq
MCF7_SEPT9_v1, MCF7_SEPT9_v2, and MCF7_SEPT9_v3-overexpressing cell lines and the MCF7 control cells (MCF7_C) were placed in culture simultaneously and split to establish three biological replicates; the plates were taken from culture when they reached 70% confluence.  The cells were then lysed directly on the plate with Qiazol lysis reagent (Qiagen, Valencia, CA) and placed at -80°C until all samples were ready for RNA extraction.  Total RNA was isolated using the miRNeasy kit (Qiagen, Valencia, CA) and assessed using an Agilent Bioanalyzer. All RNA had an RNA integrity number (RIN) score of 10. The mRNA was randomly fragmented using the RNA fragmentation kit from Ambion. First-strand cDNA synthesis was performed using random primers and SuperScriptII reverse-transcriptase (Invitrogen, Carlsbad CA). This was followed by second-strand cDNA synthesis using DNA Polymerase I and RNase H (Invitrogen, Carlsbad CA). The Illumina adaptor was ligated to the ends of the double-stranded cDNA fragments and a 200bp size-selection of the final product was performed by gel-excision, following the Illumina-recommended protocol. Template molecules of 200bp cDNA with the adaptor attached were enriched by PCR to create the final library.
GEO Accession
GSM3374813