Illumina HiSeq 2000 sequencing; BT549 cells depleted of ELP3 compared to control
BT549 cells depleted of ELP3 compared to control
BT549 cells were stably depleted of ELP3 by shRNA  a scramble shRNA was used as control.  DMEM + 10% FBS + 5% glut lentiviral infection shCTR and shRNA against ELP3 TRIZOL RNA integrity was verified on the Bioanalyser 2100 with RNA 6000 Nano chips, and RIN scores were >9 for all samples The Illumina Truseq RNA Sample Preparation kit V2 was used to prepare libraries from 500 ng total RNAs. PolyA RNAs were purified with polyT-coated magnetic beads, chemically fragmented, and used as template for cDNA synthesis using random hexamers. cDNA ends were subsequently end-blunted, adenylated at 3′OH extremities, and ligated to indexed adaptors. Finally, the adapters ligated library fragments were enriched by PCR following Illumina's protocol and purified
Experimental Factor: RNA interference
ELP3 shRNA