Mutation Quantification from Sanger Sequencing Traces
The quantitative information embedded within Sanger sequencing traces can be useful for DNA mutation, variation, and methylation analysis, including disease study. Detection and quantification of minor variants, including DNA mutation and methylation, is simplified in Mutation Surveyor software. The software rapidly and accurately aligns sample traces to a reference trace, detecting minor variants with greater than 99% accuracy and sensitivity as low as 5% of the primary peak. Mutation Surveyor software also includes a Mutation Quantifier tool, which provides a quantitative analysis of peak characteristics, enabling the user to easily calculate allelic proportions in Sanger Sequencing projects. The tool's flexibility provides two methods of quantifying peak ratios, including a simplified allele ratio based on peak Relative Fluorescence Units (RFU) and a standardized allele ratio for percent drop of the normal allele and percent gain of the mutant allele at specified locations. These locations within the genome can be quantified based on mutation calls or specified loci using reference coordinates.