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Optimized Detection of DNA Point Mutations by Double Gradient Denaturing Gradient Gel

  • Laura Cremonesi , Paola Carrera , Elena Cardillo , Antonella Fumagalli , Sabrina Lucchiari , Maurizio Ferrari , Sabina C. Righetti , Pier Giorgio Righetti and Cecilia Gelfi
Published/Copyright: July 5, 2005
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Clinical Chemistry and Laboratory Medicine (CCLM)
From the journal Volume 36 Issue 12

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.

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Published Online: 2005-07-05
Published in Print: 1998-12-05

Copyright © 1999 by Walter de Gruyter GmbH & Co. KG

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