Effects of formamide on the thermal stability of DNA duplexes on biochips.

Julia Fuchs, Daniela Dell'Atti, Arnaud Buhot, Roberto Calemczuk, Marco Mascini, Thierry Livache

UMR 5819 (UJF, CNRS, CEA) CEA, INAC, SPrAM, 38054 Grenoble cedex 9, France.

Analytical biochemistry 2010 Feb 1

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In molecular biology, formamide (FA) is a commonly used denaturing agent for DNA. Although its influence on DNA duplex stability in solution is well established, little is known about immobilized DNA on microarrays. We measured thermal denaturation curves for oligonucleotides immobilized by two standard protocols: thiol self-assembling and pyrrole electrospotting. A decrease of the DNA denaturation temperature with increasing FA fraction of the solvent was observed on sequences with mutations for both surface chemistries. The average dissociation temperature decrease was found to be -0.58+/-0.05 degrees C/% FA (v/v) independently of grafting chemistry and probe sequence. Copyright 2009 Elsevier Inc. All rights reserved.

Citation

Julia Fuchs, Daniela Dell'Atti, Arnaud Buhot, Roberto Calemczuk, Marco Mascini, Thierry Livache. Effects of formamide on the thermal stability of DNA duplexes on biochips. Analytical biochemistry. 2010 Feb 1;397(1):132-4