ANALYSIS OF SINGLE-STRAND CONFORMATION POLYMORPHISMS BY CAPILLARY ELECTROPHORESIS WITH LASER-INDUCED FLUORESCENCE DETECTION

Detection of point mutations in genomic DNA is important for diagnosis of inherited characteristics and genetic diseases. A point mutation i n a specific region of DNA amplified by polymerase chain reaction (PCR ) can be detected with single-strand conformation polymorphism (SSCP) analysis. Analysis of SSCP by laser-induced fluorescence capillary ele ctrophoresis in entangled polymer solution (CE-LIF) has been developed in the present paper. K-ras genes including seven mutations were ampl ified with primer labeled with Texas Red at its 5' end. The labeled PC R products were dissociated to single strands by heating and separated with capillary gel electrophoresis and He-Ne laser-excited fluorescen ce detection. Our results suggest that all fragments having normal (Gl y) and mutated (Ala, Arg, Cys, Ser, Val, Asp) sequences at codon 12 ca n be distinguished. Analysis of SSCPs with CE-LIF is well suited for c linical analysis of SSCPs because of its high sensitivity, resolution, reproduciblity and speed. (C) 1997 Elsevier Science B.V.