GSM6857591: T1_ENR1; Mus musculus; OTHER
DNA from all timepoints was isolated using the DNeasy Blood and Tissue Kit (Qiagen) sgRNAs were amplified from genomic DNA using KAPA HiFi HotStart Library Amplification Kit (Kapa Biosystems) with 20 cycles of PCR. A second PCR was used to add sample unique Illumina sequencing barcodes. The amplified sgRNA libraries were run in an agarose gel and isolated with Gel Extraction Kit (Qiagen) and cleaned with DNA Clean & Concentrator kit (Zymo Research). The concentrations of the sgRNA libraries were measured using KAPA Library Quantification kit (Kapa Biosystems), and diluted to 2 nM before libraries were pooled. A final dilution of 1.7 pM pooled libraries were sequenced with Illumina High Output 75 cycles kit and sequenced on Illumina NextSeq500 sequencer.