title: GSM8000757: Huh7.5.1-ACE2-TMPRSS2, uninfected, Gecko B; Homo sapiens; OTHER
library_construction_protocol: For SARS-CoV, surviving cells were harvested at 12 days post-infection in RLT buffer and genomic DNA was extracted with AllPrep DNA/RNA spin columns (Qiagen). Eluted DNA was irradiated for biosafety reasons. For MERS-CoV, surviving cells were harvested in DNA/RNA Shield (Zymo) at 8 days post-infection and genomic DNA was extracted using the Quick-DNA Midiprep Plus Kit (Zymo). The sgRNA expression cassettes were amplified from genomic DNA in a two-step nested PCR using KAPA HiFi HotStart ReadyMixPCR Kit (Kapa Biosystems) and sequenced on an Illumina NextSeq 500 using a custom sequencing primer, as previously described (PMID: 33333024). OTHER: Amplicon library of sgRNA expression cassettes from genomic DNA