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Method for the Characterization of Size-Exclusion Chromatography Media for Preparative Purification of DNA Restriction Fragments

Published

Author(s)

C M. Tellez-Rodriguez, Kenneth D. Cole

Abstract

A protocol has been developed to characterize the potential of size exclusion chromatography gels for the separation of DNA fragments at a preparative scale. A standard DNA mixture composed of Λ-DNA cut with the restriction enzymes AluI and HaeIII was chromatographed on a column packed with the gel of interest. Fractions were collected and examined by acrylamide gel electrophoresis. A digitized image of the electrophoresis gel was analyzed with the aid of a computer software program to determine the DNA fractionation range and selectivity curve for each chromatography gel. Four gels were characterized using this protocol. The effect of the elution buffer ionic strength on the fractionation of DNA was also investigated.
Citation
Biotechnology Techniques
Volume
13
Issue
6

Keywords

characterization, chromatography, digitized image, DNA, electrophoresis, parparative, restriction fragments, size exclusion

Citation

Tellez-Rodriguez, C. and Cole, K. (1999), Method for the Characterization of Size-Exclusion Chromatography Media for Preparative Purification of DNA Restriction Fragments, Biotechnology Techniques (Accessed July 20, 2024)

Issues

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Created June 1, 1999, Updated February 17, 2017