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J Vis Exp. 2010 Sep 5;(43). pii: 2136. doi: 10.3791/2136.

Electroeluting DNA fragments.

Author information

1
Department of Genetics and Molecular Biology, Research and Advanced Studies Center of National Polytechnic Institute.

Abstract

Purified DNA fragments are used for different purposes in Molecular Biology and they can be prepared by several procedures. Most of them require a previous electrophoresis of the DNA fragments in order to separate the band of interest. Then, this band is excised out from an agarose or acrylamide gel and purified by using either: binding and elution from glass or silica particles, DEAE-cellulose membranes, "crush and soak method", electroelution or very often expensive commercial purification kits. Thus, selecting a method will depend mostly of what is available in the laboratory. The electroelution procedure allows one to purify very clean DNA to be used in a large number of applications (sequencing, radiolabeling, enzymatic restriction, enzymatic modification, cloning etc). This procedure consists in placing DNA band-containing agarose or acrylamide slices into sample wells of the electroeluter, then applying current will make the DNA fragment to leave the agarose and thus be trapped in a cushion salt to be recovered later by ethanol precipitation.

PMID:
20834225
PMCID:
PMC3157863
DOI:
10.3791/2136
[Indexed for MEDLINE]
Free PMC Article

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