摘要：Genetic variations such as mutations and recombinations arise spontaneously in all cultured *** it is possible to identify nonneutral mutations by selection or counterselection,the identification of neutral mutations in a heterogeneous population usually requires expensive and time-consuming methods such as quantitative or droplet polymerase chain reaction and high-throughput *** mutations could even become dominant under changing environmental conditions enforcing transitory selection or *** propose a novel method,which we called qSanger,to quantify DNA using amplitude ratios of aligned electropherogram peaks from mixed Sanger sequencing *** expressing enhanced green fluorescent protein and mCherry fluorescent markers were used to validate qSanger both in vitro and in cotransformed Escherichia coli via quantitative polymerase chain reaction and fluorescence *** show that qSanger allows the quantification of genetic variants,including single-base natural polymorphisms or de novo mutations,from mixed Sanger sequencing reads,with substantial reduction of labor and costs compared to canonical approaches.