title: GSM8879171: day9_iNs_NT_CRISPRi_AX227_replicate2; Homo sapiens; OTHER
library_construction_protocol: For CRISPRi-screen, genomic DNA was extracted from transduced iPSCs using the DNeasy Blood & Tissue Kit (QIAGEN). The extracted DNA was then amplified by PCR to enrich target fragments for sequencing. For CRISPRi-screen, the PCR reaction included 10 μg of DNA, 1 μM of each forward and reverse primer containing Illumina sequencing indices, and 1× KAPA HiFi HotStart Ready Mix (Roche). The amplification process involved an initial 3-minute denaturation at 95°C, followed by 20 cycles of 30 seconds at 95°C, 30 seconds at 55°C, and 30 seconds at 72°C, concluding with a final 5-minute extension at 72°C. The amplified DNA was purified using AMPure XP beads (Beckman Coulter) with a 2× size selection, and the quality was assessed using the Agilent High Sensitivity D5000 ScreenTape System.
identifiers.namespace: GEO
identifiers.id: GSM8879171_r1