I used CRISPR method to make albino zebrafish and got albino(F1) by mating F0 female and male. I confirmed only 50% mutant(F2) from albino(F1) outcrossing with WT by T7E1 enzyme. The Surveyor could even not detect other 50% mutant in this case. I think it may come from point mutation. So does anyone know any simple method to find hetero point mutant except using restriction enzyme and sequencing?
You might be able to use something similar...
Article A simplified method for identifying early CRISPR-induced ind...
if you know what the mutation is then ARMS or ACRS pcr techniques or if you do not know what the change is but you do know approximately where it is then SSCP can be a useful screening technique. Sometimes if you are lucky the pcr generates hereroduplexed pcr product which can run as an extra band just slower than the normal or mutant band ( which both run at the same size) in agarose gels
Dear Paul Rutland
Thanks for kind information. In my case, the heteroduplex didn't show any difference in 4% agarose gel ( band sie 315bp, running time 80V 120min). So as your recommendation, I think SSCP analysis may be the best choice until now.
Thanks again.
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