title: GSM6971616: Screen-Experiment2-CD44-low; Homo sapiens; OTHER
library_construction_protocol: CRISPRa-sgRNA containing cells were harvested and processed for CD44+ sorting. Note, for cell sorting using BD FACSAria II sorter, 120-micron tip was used to let free flow of large mesenchymal-like cells.  We sorted ~50X representation of the library from CD44-low (bottom 10% of the peak) and CD44-high (top 10% of the peak) cells. Genomic DNA were harvested from CD44 sorted populations using the Quick-DNA plus (Zymo Research) according to the manufacturer's protocol. To determine distribution of the sgRNA, the spectrophotometer quantified gDNA product was sequenced using primers (listed in Supplementary file 1 of this paper) in Illumina MiSeq (80 cycles of read 1 (forward) and 8 cycles of index 1 with a 5% PhiX control to improve library diversity in order to cover >50 reads per sgRNA in the library).