How to design primers for PCR screening of CRIPR KO in CHO-K1 cells?

11 November 2016 0 5K Report

I have to screen CHO-K1 clones to search for KO of my gene of interest by CRISPR. For this purpose, I need to amplify the exon with the expected mutation by using primers specific of that region. How do you design region-specific primers to avoid amplification of other region within the genome? Do you think the database in primer-BLAST is a valid way to screen for off-target annelaing? Could you suggest other online database to check for the specificity of annealing in CHO-K1 genome?

Thanks

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