We are planning a gene knockout experiment using origene gene specific crisper knockout. We were wondering of the best way to select for double knockout without doing southern blot? will FISH do it or do you have a better way?
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Yuan-Yeu Yau
1. What did you mean by 'double' knockouts? You meant (1) bi-allelic knockout of the pair of homozygous chromosomes or (2) you have two genes want to be knocked out?
2. Sequencing can be one way to check whether your intended allele has been 'disrupted' due to DSB occurrence.
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Katrin Domsch
You can also try a T7 endonuclease I assay to identify which of your lines contains a mutations. And finally I would also recommend sequencing.
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