Applications of Constant Denaturant Capillary Electrophoresis and Complementary Procedures: Measurement of Point Mutational Spectra

Constant denaturant capillary electrophoresis (CDCE) separates macromolecules based on differences in their melting temperatures. The specific apparatus and operating conditions have been described previously that allow CDCE to separate point mutants among 100-150-bp iso-melting DNA sequences (1 ,2 ). CDCE coupled with high-fidelity DNA polymerase chain reaction (PCR) has been applied to the measurement of point mutational spectra in human cell and tissue samples (3 -4 ). For reviews of this field see refs. 7 ,8 . This chapter describes additional techniques which when combined with CDCE and high-fidelity PCR allow point mutation detection at fractions as low as 10⁻⁶ .