CAPILLARY ELECTROPHORESIS FOR THE DETECTION OF KNOWN POINT MUTATIONS BY SINGLE-NUCLEOTIDE PRIMER EXTENSION AND LASER-INDUCED FLUORESCENCE DETECTION

Capillary electrophoresis (CE) with laser-induced fluorescence (LIF) w as used to detect known point mutations using the method of single-nuc leotide primer extension (SNuPE). Three different point mutations in h uman mitochondrial DNA associated with Leber's hereditary optic neurop athy (LHON) were detected by annealing a primer immediately 5' to the mutation on the template and extending the primer by one fluorescently labeled dideoxy terminator complementary to the mutation. By using tw o or more differently labeled terminators, both the mutant and wild ty pe could be simultaneously detected. The advantages of using CE-LIF fo r detecting SNuPE reactions include speed and ease of analysis, absenc e of radioactivity, and potential for automation. (C) 1997 Elsevier Sc ience B.V.