Figure 1 | Laboratory Investigation

Figure 1

From: A simple method based on Sanger sequencing and MS Word wildcard searching to identify Cas9-induced frameshift mutations

Figure 1

Overview of SWS strategy design. (a) A schematic graph showing the patterns of indels induced by CRISPR/Cas9 system. sgRNAs target site (yellow); PAM (5’NGG3’, pink); cleavage site cut (red triangle); indels (red bars). CUIS stands for the common DNA sequences upstream of indels, which are identical to corresponding wild-type genomic sequences and shared by both alleles (green bars). CDIS is the common DNA sequences downstream of indels, which are identical to corresponding wild-type genomic sequences and shared by both alleles (blue bars). (b) A graph showing the analysis of Sanger sequencing graph. The Sanger sequencing graph was made by MS drawing software. Green single peaks indicate CUIS; red double peaks indicate putative indels; blue double peaks indicate CDIS. A 7- to 10-bp length CF (N[N/N][N/N][N/N][N/N]N[N/N]) 5- to 100-bp (n) downstream of CUIS is selected for wildcard searching. (c) A screenshot showing MS Word wildcards searching of CF (N[N/N][N/N][N/N][N/N]N[N/N]) in wild-type sequence. (d) A schematic graph showing the identification of corresponding CF′s (CF′1 and CF′2) in wild-type sequence and the calculation of the length of indels.

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