Multiplex, fluorescent single-strand conformation polymorphism using stepped polymerase chain reaction primers.

A method for high-throughput screening for a well-characterized mutation at the APC locus was developed using stepped PCR primers, pooled product, and fluorescent single-strand conformation polymorphism (SSCP). Using a panel of two known normals, three known heterozygous mutants, and one known homozygous mutant, eight stepped PCR products were developed that showed the same mobility shift on a PE Biosystems 373A. Using this method, up to eight different samples can be pooled for same lane analysis.