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Rapid classification of phenylketonuria genotypes by analysis of heteroduplexes generated by PCR‐amplifiable synthetic DNA
Author(s) -
Wood Nigel,
Tyfield Linda,
Bidwell Jeffrey
Publication year - 1993
Publication title -
human mutation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 162
eISSN - 1098-1004
pISSN - 1059-7794
DOI - 10.1002/humu.1380020213
Subject(s) - biology , heteroduplex , genotyping , microbiology and biotechnology , exon , genetics , phenylalanine hydroxylase , point mutation , genotype , polymerase chain reaction , mutation , dna , mutant , gene , phenylalanine , amino acid
We describe a rapid and simple method for phenylketonuria genotyping which identifies five point mutations within exon 12 of the human phenylalanine hydroxylase gene. The method involves PCR amplification of the target exon and hybridization with a PCR‐amplifiable synthetic DNA (universal heteroduplex generator, UHG). The UHG contains identifiers consisting of nucleotide substitutions and/or deletions, contiguous with known mutation sites within the target exon. DNA heteroduplexes are resolved by nondenaturing polyacrylamide minigel electrophoresis. Individual mutant genotypes are identified by characteristic banding patterns, in either homozygous or heterozygous states. The method may potentially be applied to rapid genotyping of any mutation or series of mutations within PCR‐amplifiable genetic material. © 1993 Wiley‐Liss, Inc.