A rapid one-tube PCR method for simultaneously differentiating homozygotes and heterozygotes of the Sp1 binding site polymorphism in collagen type Ialpha1.
Rapid detection of single-base changes is fundamental to molecular medicine. PCR amplification of specific alleles (PASA) has previously been used as a rapid method of genotyping single-nucleotide changes, but one reaction is required for each allele. This paper describes a Bidirectional PASA (Bi-PASA) method, which was developed to distinguish between homozygotes and heterozygotes in one PCR reaction. The method is tested using the Sp1 polymorphism in Collagen type Ialpha1. The results demonstrate that Bi-PASA is a simple and rapid method for detecting the zygosity of the polymorphism in a single PCR reaction.
Lundberg
Gunilla A
GA
Department of Clinical Medicine, Orebro University, S-701 82 Orebro, Sweden. gunilla.lundberg@ikm.oru.se
D001483:Base Sequence; D001665:Binding Sites; D024042:Collagen Type I / Q000235:genetics*; D000090602:Collagen Type I, alpha 1 Chain; D004252:DNA Mutational Analysis / Q000379:methods; D006579:Heterozygote; D006720:Homozygote; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D011110:Polymorphism, Genetic; D016329:Sp1 Transcription Factor / Q000378:metabolism*
D024042:Collagen Type I; D000090602:Collagen Type I, alpha 1 Chain; D016329:Sp1 Transcription Factor