Amplicon sequencing of Cdc42 locus in PLB-985 Cdc42 KO cell line
Genomic DNA was harvested from 5 million cells (Invitrogen Purelink Genomic DNA Kit). The purified gDNA was then used as a PCR template for primers that flank the CRISPR cut site. The forward primer sequence was 5-ACACTCTTTCCCTACACGACGCTCTTCCGATCTccagcatgcttttaacactttgagg, and the reverse primer sequence was 5-GACTGGAGTTCAGACGTGTGCTCTTCCGATCTgaaaggagtctttggacagtggtg. The PCR product was cleaned (Zymo DNA clean and concentrator-5) and then sent to Genewiz for 2 x 250 bp amplicon sequencing (Amplicon-EZ service)