sgRNA for SDHC was designed targeting the intron region just before the Exon 1 of sdhc.“CACCG” was added towards the 5′ end of 20-nt guide sequence. “AAAC” was added at the 5′ end and an extra “C” towards the 3′ end of the opposite pair of sgRNA. sgRNA pairs were ...
were used to amplify the cDNA sequences isolated from the CRISPR-edited cells. PCR products were electrophoresed in a 1% agarose gel and the gel bands were isolated using QIA Quick PCR Purification Kit (Qiagen). Isolated DNA was cloned into pCR-TOPO plasmids using the TOPO TA cloning kit ...