GSM9194694: iPSC-Derived Microglia CRISPRi Screen, d12; Homo sapiens; OTHER
CRISPRi screen cell extraction: Cells were collected and centrifuged, resuspended in PBS and subjected to DNA extraction (Qiagen DNeasy) per protocol at days 0, 12, 24, 36 and 45 of differentiation. CRISPRi NGS library construction: Genomic DNA was amplified with barcoded NGS primers, and reactions were pooled and concentrated (Zymo). Libraries were size selected using SPRI beads with dual selection to enrich for 275bp amplicons. Libraries were spiked with PhiX and prepared for sequencing with Illumina iSeq 100.