Optimized detection of DNA point mutations by double gradient denaturing gradient gel electrophoresis

L. Cremonesi, P. Carrera, E. Cardillo, A. Fumagalli, S. Lucchiari, M. Ferrari, S. C. Righetti, P. G. Righetti, C. Gelfi

Research output: Contribution to journalArticle

Abstract

Denaturing gradient gel electrophoresis displays the highest detection rate among mutation scanning methods. In classical denaturing gradient gel electrophoresis the denaturant gradient range and migration times vary for every amplicon to be scanned, greatly affecting the routine application of the method. As an alternative, we developed double gradient denaturing gradient gel electrophoresis where a gradient of pore size is superimposed over the denaturing one, allowing maintenance of the zone-sharpening effect even over prolonged time runs, and adoption of identical run time conditions for all fragments analyzed. Here double gradient denaturing gradient gel electrophoresis has been applied to the analysis of a number of point mutations and polymorphisms located in several exons of three different genes, the cystic fibrosis transmembrane conductance regulator, the β-globin and the p53 genes.

Original languageEnglish
Pages (from-to)959-961
Number of pages3
JournalClinical Chemistry and Laboratory Medicine
Volume36
Issue number12
DOIs
Publication statusPublished - 1998

Keywords

  • Denaturing gradient gel electrophoresis (DGGE)
  • Double gradient denaturing gradient gel electrophoresis (DG-DGGE)
  • Mutations
  • Scanning methods

ASJC Scopus subject areas

  • Clinical Biochemistry

Fingerprint Dive into the research topics of 'Optimized detection of DNA point mutations by double gradient denaturing gradient gel electrophoresis'. Together they form a unique fingerprint.

  • Cite this