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Nucleic Acids Research, Vol 25, Issue 15 3059-3065, Copyright © 1997 by Oxford University Press

ARTICLES

Modification of DNA duplexes to smooth their thermal stability independently of their base content for DNA sequencing by hybridization

HK Nguyen, P Auffray, U Asseline, D Dupret and NT Thuong
Centre de Biophysique Moleculaire, CNRS, rue Charles Sadron, 45071 Orleans Cedex 02, France and 1 Appligene-Oncor, Parc d'Innovation, BP 72, 67407 Illkirch, France.

The possibility of equalizing DNA duplex stability is essential for the application of sequencing by hybridization. In this paper we describe a new strategy to obtain DNA duplexes with a thermal stability independent of their base content. Modified *C bases have been developed and incorporated into oligonucleotides. The influence of these modifications on duplex stability has been studied by absorption spectroscopy, thus allowing selection of N -4-ethyl-2'-deoxycytidine (d4EtC), which hybridizes specifically with natural dG to give a G4EtC base pair whose stability is very close to that of natural AT base pairs. Duplexes built with AT and/or G4EtC base pairs exhibit thermal stabilities independent of their base content in a classical buffer solution, thus enabling control of the stability of DNA hybrids as a function of their length only.
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