Identification of DNA Polymorphism by Asymmetric-PCR SSCP

摘要

Single-strand conformation polymorphism combined with PCR (PCR-SSCP) has become an important tool for gene analysis, particularly in detecting point mutations (8) and typing of DNA polymorphisms (3,6). Although this method has sufficient sensitivity to detect single-base substitutions in many cases, there are still some problems to overcome in applying SSCP to DNA polymorphism analysis. One is that a relatively high resolution is requited because polymorphic regions normally have so many DNA types that their SSCP patterns tend to be more complicated, especially when the het-erozygotes are examined. A second problem is that SSCP gel electrophore-sis needs reference samples. Since typing is carried out by matching mobility patterns with reference samples, as many reference lanes as there are DNA types are needed.