A process for the coupling of sscp and dgge for improved recognition of point mutations

摘要

A mutation screening method (I), is new and comprises the coupling of single strand conformation polymorphism (SSCP) and denaturing gradient gel electrophoresis (DGGE) through the use of a nested polymerase chain reaction (PCR) primer with a GC adapter sequence. A mutation screening method (I), is new and comprises the coupling of single strand conformation polymorphism (SSCP) and denaturing gradient gel electrophoresis (DGGE) through the use of a nested polymerase chain reaction (PCR) primer with a GC adapter sequence, using: (1) SSCP analysis of the nested PCR products (with GC adapters) of all samples; (2) DGGE mutation analysis of the nested PCR products (with GC adapters) plus attached universal GC clamps of all samples showing no mutation signal after SSCP analysis; (3) sequence analysis of the mutations detected by one of the two screening methods; and optionally (4) post-amplification using a universal biotin-labeled sequence primer.