Genotyping method for CRISPR/Cas9 Knockout mouse

Gene Summary

Gene Name

Model Description

For knockout mice obtained by random repair via NHEJ, their genotypes can generally be determined by PCR+ sequencing.See the following figure for details: <img src="default/20180820/f9e96decc2c11e1278f0c5b9b0b3d16f.jpg" title="2-xg.jpg" alt="2-xg.jpg" class="img-responsive"> <p style="text-align: justify;">Figure 9. 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, a number of different genotypes can be generated due to repair by NHEJ, thus producing noise peaks near the digestion point of Cas9. 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 (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 C" loses two bases compared to C'. The peak shapes of C' and C" are consistent with that of C. 
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