Silver Staining of Denaturing Gradient Gel Electrophoresis Gels

摘要

The continuing progress in the identification and characterization of genes that cause human genetic diseases has greatly increased the requests for a method that can rapidly identify mutations in these genes. The introduction of PCR has made it possible to rapidly obtain a relatively large amount of single-copy genomic DNA that can be used for subsequent analysis. Direct sequencing of amplified DNA, although a standard procedure in many laboratories, is still time-consuming and laborious. Consequently, different methods have been developed to pre-identify the region of the gene in which the mutation is located, which can subsequently be amplified and sequenced. We have chosen denaturing gradient gel electrophoresis (DGGE) of PCR-amplified material, by which fragments up to 800 bp in length can be screened for the presence of mutations. DGGE, as described originally (1), relies …