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Electroeluting DNA Fragments

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...

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Autores principales: Zarzosa-Álvarez, Ana L., Sandoval-Cabrera, Antonio, Torres-Huerta, Ana L., Ma. Bermudez-Cruz, Rosa
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MyJove Corporation 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3157863/
https://www.ncbi.nlm.nih.gov/pubmed/20834225
http://dx.doi.org/10.3791/2136
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author Zarzosa-Álvarez, Ana L.
Sandoval-Cabrera, Antonio
Torres-Huerta, Ana L.
Ma. Bermudez-Cruz, Rosa
author_facet Zarzosa-Álvarez, Ana L.
Sandoval-Cabrera, Antonio
Torres-Huerta, Ana L.
Ma. Bermudez-Cruz, Rosa
author_sort Zarzosa-Álvarez, Ana L.
collection PubMed
description 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.
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spelling pubmed-31578632011-11-15 Electroeluting DNA Fragments Zarzosa-Álvarez, Ana L. Sandoval-Cabrera, Antonio Torres-Huerta, Ana L. Ma. Bermudez-Cruz, Rosa J Vis Exp Basic Protocols 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. MyJove Corporation 2010-09-05 /pmc/articles/PMC3157863/ /pubmed/20834225 http://dx.doi.org/10.3791/2136 Text en Copyright © 2010, Journal of Visualized Experiments http://creativecommons.org/licenses/by-nc-nd/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. To view a copy of this license, visithttp://creativecommons.org/licenses/by-nc-nd/3.0/
spellingShingle Basic Protocols
Zarzosa-Álvarez, Ana L.
Sandoval-Cabrera, Antonio
Torres-Huerta, Ana L.
Ma. Bermudez-Cruz, Rosa
Electroeluting DNA Fragments
title Electroeluting DNA Fragments
title_full Electroeluting DNA Fragments
title_fullStr Electroeluting DNA Fragments
title_full_unstemmed Electroeluting DNA Fragments
title_short Electroeluting DNA Fragments
title_sort electroeluting dna fragments
topic Basic Protocols
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3157863/
https://www.ncbi.nlm.nih.gov/pubmed/20834225
http://dx.doi.org/10.3791/2136
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