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Updated: Aug 23, 2026

Iterative Optimization of DNA Duplexes for Crystallization of SeqA-DNA Complexes
Published on: November 1, 2012
Modification of DNA duplexes to smooth their thermal stability independently of their base content for DNA sequencing
H K Nguyen1, P Auffray, U Asseline
1Centre de Biophysique Moléculaire, CNRS, rue Charles Sadron, 45071 Orléans Cedex 02, France and 1 Appligène-Oncor, Parc d'Innovation, BP 72, 67407 Illkirch, France.
Abstract:
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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