Small alterations in DNA sequences of genes lead to many human diseases, such as cancer, diabetes, heart disease, myocardial infarction, atherosclerosis, cystic fibrosis, and Alzheimer’s disease. These alterations in DNA sequence include many types of mutations and polymorphisms, such as nucleotide substitutions, deletion or insertion of some sequence, differences in a variable number of tandem repeat (VNTR) locus, and instability of microsatellite repeat sequences.
CGE is a particularly suitable method for carrying out analysis of DNA fragments for diagnostic purposes. Currently available methods for analysis of mutations and polymorphisms include some modified polymerase chain reaction (PCR) techniques, restriction fragment length polymorphism (RFLP), single-strand conformation polymorphism (SSCP), VNTR, and short tandem repeats (STRs) analysis, and hybridization techniques, as well as PCR analysis itself.
Labeled DNA fragments to be analyzed by CE are obtained by the incorporation of fluorescent-dye-labeled nucleotides or fluorescent dye-labeled primers during the PCR reaction. The range of available dyes and the reduction in costs over recent years have allowed modification of existing tests to take advantage of the four/five-color detection system and the degree of automation offered by commercially available CE electrophoresis systems.
Four-color, highresolution analysis is particularly useful for the rapid mapping of genetic traits, leading to gene discovery and eventual diagnostic testing. Automation is essential for rapid gene-based mutation analysis in clinical laboratories to screen a large number of DNA samples.
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