Figure 5
Illustration of heteroduplex DNA formation from plasmids harboring various indel mutations by PAGE analysis.
(A) Sequence analysis identified DNA plasmids from Them2 locus harboring -2 bp, -3 bp, -4 bp, -6 bp, -7 bp, -8 bp and -16 bp genomic deletion. (B-C) Agarose gel (B) and PAGE-based approach (C) for detection of various heteroduplex DNA. Agarose gel electrophoresis failed to detect indel mutations when mixing plasmids from wildtype allele with plasmids harboring various indel mutations; PAGE analysis revealed heteroduplex DNA formation as expected. (D) Schematic overview of heteroduplex DNA formation in case of samples harboring biallelic indel mutations. Red and blue boxes represent two type of indel mutations. After denaturation and annealing of PCR products, three types of homoduplex DNA and six types of heteroduplex DNA were formed. (E–F) Agarose gel (E) and PAGE-based approach (F) for detection of various combinations of heteroduplex DNA. PAGE analysis revealed heteroduplex DNA formation when mixing plasmids harboring wildtype allele with plasmids harboring two types of indel mutations, whereas agarose gel electrophoresis failed to detect any indel mutation.
