Variability in the interpretation of microsatellite patterns with different electrophoretic conditions
Document Type
Journal Article
Publication Date
1-1-1999
Journal
Journal of Clinical Pathology - Molecular Pathology
Volume
52
Issue
5
DOI
10.1136/mp.52.5.302
Keywords
Electrophoresis; Interpretation; Microsatellite instability
Abstract
Microsatellite markers permit the analysis of microsatellite instability and loss of heterozygosity. Frequently, the allelotypes of microsatellites are interpreted in the presence of numerous bands in gels. The importance of different gel electrophoresis conditions in the interpretation of microsatellite patterns was tested. Microsatellite markers were used to amplify DNA from gastric cancer samples and adjacent gastric mucosa. Polymerase chain reaction (PCR) products were separated by electrophoresis through 7% polyacrylamide gels containing either 5.6 M urea and 32% formamide or 7 M urea. PCR reactions separated on urea/formamide gels resulted consistently in clear allele definition (one or two bands), whereas 7 M urea gels resulted in allele patterns that comprised multiple bands. Analysis of microsatellite abnormalities using nonformamide gels gave false negative results in just under a third of cases (four of 13). In conclusion, the interpretation of microsatellite alterations in cancer DNA is improved by using electrophoresis conditions that result in complete DNA denaturation, such as urea/formamide/acrylamide gel electrophoresis.
APA Citation
Santos, A., Yamaoka, Y., Graham, D., & Sepulveda, A. (1999). Variability in the interpretation of microsatellite patterns with different electrophoretic conditions. Journal of Clinical Pathology - Molecular Pathology, 52 (5). http://dx.doi.org/10.1136/mp.52.5.302