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ssDNA degradation along capillary electrophoresis process using a Tris buffer

Ric, Audrey and Ong-Meang, Varravaddheay and Poinsot, Verena and Martins-Froment, Nathalie and Chauvet, Fabien and Boutonnet, Audrey and Ginot, Frédéric and Ecochard, Vincent and Paquereau, Laurent and Couderc, François ssDNA degradation along capillary electrophoresis process using a Tris buffer. (2017) ELECTROPHORESIS, 38 (12). 1624-1631. ISSN 0173-0835

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Official URL: https://doi.org/10.1002/elps.201600561

Abstract

Tris-Acetate buffer is currently used in the selection and the characterization of ssDNA by capillary electrophoresis (CE). By applying high voltage, the migration of ionic species into the capillary generates a current that induces water electrolysis. This phenomenon is followed by the modification of the pH and the production of Tris derivatives. By injecting ten times by capillary electrophoresis ssDNA (50 nM), the whole oligonucleotide was degraded. In this paper, we will show that the Tris buffer in the running vials is modified along the electrophoretic process by electrochemical reactions. We also observed that the composition of the metal ions changes in the running buffer vials. This phenomenon, never described in CE, is important for fluorescent ssDNA analysis using Tris buffer. The oligonucleotides are degraded by electrochemically synthesized species (present in the running Tris vials) until it disappears, even if the separation buffer in the capillary is clean. To address these issues, we propose to use a sodium phosphate buffer that we demonstrate to be electrochemically inactive.

Item Type:Article
HAL Id:hal-02134843
Audience (journal):International peer-reviewed journal
Uncontrolled Keywords:
Institution:French research institutions > Centre National de la Recherche Scientifique - CNRS (FRANCE)
Université de Toulouse > Institut National Polytechnique de Toulouse - INPT (FRANCE)
French research institutions > Institut National de la Recherche Agronomique - INRA (FRANCE)
French research institutions > Institut de Recherche pour le Développement - IRD (FRANCE)
Université de Toulouse > Université Toulouse III - Paul Sabatier - UPS (FRANCE)
Other partners > Picometrics Technologies (FRANCE)
Laboratory name:
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Deposited By: Loetitia MOYA
Deposited On:08 Oct 2018 16:24

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