20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
connexin 32 CRISPR/Cas9 Knockout (KO) Plasmid (m) is a pool of plasmids, each encoding Cas9 nuclease and a target-specific 20 nt guide RNA (gRNA) designed for maximum knockout efficiency using sequences derived from the GeCKO v2 library
gRNA sequences direct Cas9 to induce site-specific double-strand breaks (DSBs) in the connexin 32 genomic locus, resulting in gene knockout through non-homologous end joining (NHEJ)
connexin 32 HDR Plasmid (m) (sc-420564-HDR) is recommended for co-transfection with connexin 32 CRISPR/Cas9 KO Plasmid (m) to enable selection of successfully edited cells through HDR-mediated integration of a puromycin resistance cassette and RFP reporter gene
connexin 32 HDR Plasmid (m) is a pool of plasmids, each containing a homology-directed repair (HDR) template corresponding to the gRNA target sites in the connexin 32 CRISPR/Cas9 KO Plasmid (m)
Each HDR plasmid contains two ~800 bp homology arms flanking the puromycin resistance and RFP cassettes, designed to bind genomic DNA sequences surrounding the Cas9-induced double-strand break site and facilitate precise HDR-mediated integration
The puromycin resistance and RFP genes are flanked by LoxP sites, enabling removal of selection markers via Cre recombinase (Cre Vector: sc-418923) after establishing stable knockout cell lines
Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: connexin 32 Antibody (HAM8): sc-21794