Abstract: A process for distinguishing nucleic acid segments on the basis of nucleotide differences, thereby providing a rapid, convenient means for detecting mutation, is disclosed. The present process comprises synthesizing separately complementary nucleic acid strands on each of at least two target nucleic acid templates using a nucleic acid polymerase and nucleoside triphosphate substrates, wherein at least one of the natural nucleoside triphosphate substrates is replaced with a mobility-shifting analog; denaturing the synthesized strands from the templates if necessary; and comparing the mobility of the separately synthesized strands through a size-fractionation medium. A difference in mobility indicates that the target nucleic acid templates contain different numbers of total nucleotides and/or different numbers of nucleotides complementary to the mobility-shifting analogs.