Genotyping method for CRISPR/Cas9 point mutation mouse

Gene Summary

Gene Name

Model Description

Genotyping is generally performed by PCR + sequencing methods. See the following figure for details: <img src="default/20180820/747005c20c31f86b562c623065f2f46f.jpg" title="1.jpg" alt="1.jpg" class="img-responsive"> <p style="text-align: justify;">图10. A.PCR products sequencing chromatogram of wild-type mouse.B. PCR products sequencing chromatogram of F0 generation positive mice.After the action of CRISPR/Cas9, repair via non-homologous end joining (NHEJ) may also occur along with the repair via homologous recombination (HR).Therefore, noise peaks may occur near the point of digestion.C. PCR products sequencing chromatogram of F1 generation positive mice.In the genome of F1 generation positive mice, one copy is a wild type and the other copy is a mutant. The sequencing of the region in the vicinity of the Cas9 action site shows two peaks near the site of mutation (two peak shapes are produced for the wild type and mutant genotypes, one for each genotype). C' and C" are the results of monoclonal sequencing of PCR products in Figure C. C' is a wild type and C‘' is a mutant. The comparison of peak shapes reveals that the mutation in C" is GTT, compared to the CCT mutation at the target mutation site in C', while the peak shapes of C' and C" are consistent with that of C.
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