GSM8060256: Long-read sequencing of COL7A1 on-target site on sample treated with Cas9 nuclease + ssODN g1; Homo sapiens; OTHER
Genomic DNA was extracted using the ReliaPrep™ Blood gDNA Miniprep System kit (Promega) according to the manufacturer's protocol A 5 kb amplicon spanning the COL7A1, COL17A1 or LAMA3 target site was amplified from 200 ng of genomic DNA by a 30-cycle PCR using KAPA HiFi HotStart PCR Kit (Roche). PCR fragments were purified using AMPure XP beads (Beckman Coulter) at a 1.8 bead to sample ratio. Libraries were prepared using the Nanopore Native Barcoding Kit 24 V14 (SQK-NBD114.24) according the manufacturer's protocol and sequenced on a MinION Flow Cell. Libraries were prepared using the Nanopore Native Barcoding Kit 24 V14 (SQK-NBD114.24) according the manufacturer's protocol and sequenced on a MinION Flow Cell.