Point mutations, the mismatch of one nucleotide base pair, are the most common type of mutations. These point mutations, if found on several important parts of a gene, may have a
serious impact. For example, the growth of human tumors have been associated with point mutations in the p53 gene (1). Past surveys have used loss of heterozygosity as an indirect essay for
inactivation of genes (2). However, although this method gives rapid results, it cannot detect point mutations. The most sensitive screening is to sequence the actual genome, but this method is
requires intensive time and labor. More recently, there has been several nucleic acid-based screening methods that can detect mutations within short fragments of DNA, including denaturing gradient
gel electrophoresis (DGGE) (3). Constant denaturant gel electrophoresis (CDGE) (4) is a modification of DGGE. The separation principle of CDGE is based on the melting behavior of double-stranded
DNA. DNA strands with mismatches are destabilized by the mismatch, since the hydrogen bonds between mismatched base pairs are weaker, and therefore is unstranded (or “melts”) at a lower temperature
than strands without mismatches. Strand separation can be detected as a reduction in the mobility of the fragment as it moves through an acrylamide gel containing a chemical denaturant. In this
study, the effectiveness of CDGE is investigated with DNA fragments of known sequences.
Publisher
Cornell Center for Materials Research
Date
2005-08-17
Copyright Notice
Copyright 2006 CCMR. Materials from the CCMR website may not be used without permisson.
Copyright Agreement
on
Additional Notes
Acknowledgements: This work was supported by the Cornell Center for Materials Research and the National Science Foundation
Guest - January 07, 2009
View Generic Document: Screening of Point Mutation in DNA Using Constant Denaturant Gel Electrophoresis 