title: GSM7795465: rep1_myh9_facs_low; Homo sapiens; OTHER
library_construction_protocol: Cells were counted following sorting to verify enrichment, followed by DNA recovery/extraction from fixed cells using the PicoPure DNA extraction kit (ThermoFisher) according to manufacturer's instructions. Recovery digests were performed for 20 hours at 65C using up to 1.5M HFF cells per reaction volume. All gDNA was split between sample-indexed 100uL Q5 PCR reactions (up to ~340ng max input per 100uL reaction) to amplify out the gRNA protospacer from HFF cells. These PCRs from gDNA were run as follows [ 98C for 30s / 25x: 98C for 10s, 60C for 30s, 72C for 15s / 72C for 2 min] with primers in Supplementary Table 15, followed by individual PCRs being pooled together and subjected to a double-sided 0.65X/1X SPRI clean-up. Quality control was performed by running the amplified libraries out on a High Sensitivity D1000 Tapestation (Agilent) to confirm expected size, and Qubit dsDNA HS assays were performed to determine a final concentration.