Single strand conformation polymorphism (SSCP) パターンに及ぼす泳動条件の影響

We investigated the effects of electrophoretic gels, buffers or temperature on SSCP pattern by an automated electrophoretic system, Phast System. The electrophoresis on the 12.5% homogeneous gel gave the greatest difference in the mobility shift by genetic mutation. The SDS-containing buffer increased the resolution of each band as well as the sensitivity of detection in most cases. However, for certain mutated sequences, the native buffer was superior at detecting mutations. The mobility shift is also temperature dependent irrespective of native or SDS buffer condition. In any new mutation sequences, it is difficult to determine which is the better condition to detect the mutations, SDS or native buffer; however, the SDS-containing buffer may lead to a better sensitivity of detection. In conclusion, it is important to perform SSCP analysis under SDS buffer condition as well as native buffer condition in all cases