GSM6593509: MCF10A, TGF-beta dosage effect; Homo sapiens; RNA-Seq
MCF10A cells were first labelled with Perturb-seq vectors without sgRNA expression using guide barcodes (GBCs) that were originally used to identify sgRNAs. Barcoded MCF10A cells were then treated with different dosages of TGF-β for 14 days and single cells were prepared and mixed at a concentration of approximately 1,000 cell/μL in ice-cold PBS. GBC to Concentration (pM) conversion: GBC1: 0 GBC2: 12.5 GBC3: 25 GBC4: 50 GBC8: 100 GBC9: 200 GBC12: 400 GBC13: 800 RNA-Seq library was prepared using 10x Chromium Single Cell 3' –end Reagent Kits User Guide (v2 Chemistry) Libraries were sequenced using paired-end sequencing (26bp Read 1 and 98bp Read 2) with a single sample index (8bp) on the Illumina HiSeq 2500