Submerged gel electrophoresis on Spreadex gels - a new method for APC genemutation detection

Familial adenomatous polyposis (FAP) is an autosomal dominant inherited dis ease. Patients with FAP develop hundreds to thousands of adenomatous polyps in the colon and rectum during their 2nd or 3rd decades, and one or more o f them progress to cancer if left without surgical treatment. The gene resp onsible for FAP was identified in 1991 and termed the APC (adenomatous poly posis coli) gene. Following identification of APC, a number of germ-line mu tations responsible for the development of the disease were found. The purp ose of this study was to determine the usefulness of a new method, submerge d gel electrophoresis, in the detection of the most-frequent mutation of th e APC gene [5-base pair (bp) deletion in codon 1309], especially in the pre symptomatic diagnosis of FAR Genomic DNAs were isolated from peripheral blo od of patients and their relatives. We used two methods, electrophoresis on polyacrylamide gel and submerged gel electrophoresis, for the identificati on of APC gene codon 1309 mutation. After only 110 min PCR fragments of 91 bp and 86 bp (5-bp deletion) were completely resolved on a Spreadex EL300 g el. Our results showed that electrophoresis using Spreadex gels provides a simple and rapid non-radioactive method for determination of the most-frequ ent germ-line mutations in the APC gene.