GSM6171876: Donor5 gRNA2 AmpSeq; Homo sapiens; OTHER
~100,000 cells per condition were resuspended in 20 μl of Quickextract DNA Extraction Solution (Epicenter) based on manufacturer recommendations. 1 μl of QE mixture was added to a two-step PCR amplicon sequencing approach using NEB Q5 2X 5 2X Master Mix Hot Start Polymerase with the manufacturer's recommended thermocycler conditions. After amplicons of about 200 bp in length were generated, a 1.0X SPRI purification was used before samples a second PCR was then used to append P5 and P7 illumina sequencing adaptors and donor-specific barcodes to amplicons. The second PCR was following by a 1.0x SPRI purification before sequencing. Amplicon sequencing