Rapid heteroduplex analysis by capillary electrophoresis.
The very important parameters for mutation screening are short time of analysis and high throughput. The analytical platform which fulfills these criteria most satisfactorily is capillary electrophoresis. Here we show the influence of several parameters such as temperature, presence of glycerol, capillary length and polymer concentration on the electrophoretic properties of DNA duplexes and evaluate their contribution to the overall time of analysis. The careful optimization of analyzed conditions allowed us to significantly decrease the time required for the detection of the 185delAG and 4153delA mutations by heteroduplex analysis. It enabled us to analyze these typical BRCA1 gene deleterious mutations in several minutes only by using very popular and widely accessible capillary electrophoresis instrumentation.
Kozlowski
Piotr
P
Laboratory of Cancer Genetics, Institute of Bioorganic Chemistry, Polish Academy of Sciences, Noskowskiego 12/14, 61-704 Poznan, Poland.
Olejniczak
Marta
M
Krzyzosiak
Wlodzimierz J
WJ
D019075:Electrophoresis, Capillary / Q000379:methods*; D019398:Genes, BRCA1; D006801:Humans; D009154:Mutation; D009692:Nucleic Acid Heteroduplexes
D009692:Nucleic Acid Heteroduplexes