BT20_24hr_DMSOvERLOTINIB_RNAseq and WHOLE GENOME CRISPR screen

BT20 (TNBC) cells were cultured with either DMSO (v/v%) or Erlotinib (10uM) for 24 hours to observe differential expression changes between conditions. CAS-9 expressing BT20 (TNBC) cells were transduced with a whole-genome library of CRISPR sgRNAs; then, cultured them with either DMSO (v/v%) or Erlotinib (10uM) and collected samples to observe differential sgRNA abundances between conditions. Overall design: BT20 cells were cultured appropriately and treated for 24 hours --> Phenol-Chloroform extraction of RNA --> TruSeq HT Stranded (mRNA) library preparation --> Whole library sequenced on Illumina Nextseq platform BT20 cells were cultured appropriately and treated for 3-4 population doublings --> gDNA was isolated using phenol-chloroform extraction --> 2 PCRs were done to a) amplify sgRNAs and b) to attach barcodes for library preparation --> Whole library sequenced on Illumina Nextseq platform