HUMAN MUTATIONS DETECTED BY GC-CLAMPED DENATURING GRADIENT GEL-ELECTROPHORESIS

The detection and characterization of single nucleotide changes in gen omic DNA is still a technical problem in the genetic analysis of human inherited disorders. There are limited possibilities of simple and ef fective methods of mutation detection, due also to the large size of t he genes to be screened. The direct detection of mutations and the ide ntification of polymorphic changes in genomic sequences within or flan king: genes are the two main applications of experimental approach. De naturing gradient gel electrophoresis (DGGE) allows a rapid screening for single base changes in enzymatically amplified DNA, i.e. by the us e of polymerase chain reaction (PCR). The method is based on the migra tion of double-stranded DNA molecules through polyacrylamide gels cont aining linearly increasing concentrations of denaturing agent.