GSM6448370: HNP SCZ MPRA DNA; Adeno-associated virus; OTHER
RNA was extracted from each well using Qiagen RNeasy kit (Qiagen, cat#74004), using 10 uL of Mercaptoethanol (Sigma-Aldrich, cat#60-24-2) per 1 mL of Qiagen RLT buffer. The columns were treated with DNase (Qiagen, cat#79256). To acquire an initial input of DNA put into the cells, DNA was extracted from the AAV which contained the AAV-MPRA library using a NucleoSpin virus kit (Macherey-Nagel, cat#740983.50). The variant and barcode region of the AAV-variant-barcode library was PCR amplified using NEBNext 2X Q5 Hifi HS Mastermix with primers that contain Illumina P5 and P7 adapters (Bcmap_P5_AAV_R and Bcmap_P7_AAV_F). The PCR product was cleaned up using Zymo PCR clean and concentrator-5. The resulting library was sequenced using custom sequencing primers (BCmap_R1Seq_AAV_R and BCmap_R2Seq_AAV_F) via Novaseq 6000 SP (2x250bp) by the UNC High-Throughput Sequencing Facility (HTSF).