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Eliminating blurry bands in gels with a simple cost-effective repair to the gel cassette

Gel electrophoresis and subsequent imaging using phosphorimagers is one of the most important and widely used techniques in RNA and DNA analysis. Radiolabeling nucleic acids with (32)P and detecting bands using a phoshorimager are useful both in a qualitative sense for nucleic acid detection and in...

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Autores principales: Bingaman, Jamie L., Frankel, Erica A., Hull, Chelsea M., Leamy, Kathleen A., Messina, Kyle J., Mitchell, David, Park, Hongmarn, Ritchey, Laura E., Babitzke, Paul, Bevilacqua, Philip C.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory Press 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5113212/
https://www.ncbi.nlm.nih.gov/pubmed/27852929
http://dx.doi.org/10.1261/rna.059303.116
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author Bingaman, Jamie L.
Frankel, Erica A.
Hull, Chelsea M.
Leamy, Kathleen A.
Messina, Kyle J.
Mitchell, David
Park, Hongmarn
Ritchey, Laura E.
Babitzke, Paul
Bevilacqua, Philip C.
author_facet Bingaman, Jamie L.
Frankel, Erica A.
Hull, Chelsea M.
Leamy, Kathleen A.
Messina, Kyle J.
Mitchell, David
Park, Hongmarn
Ritchey, Laura E.
Babitzke, Paul
Bevilacqua, Philip C.
author_sort Bingaman, Jamie L.
collection PubMed
description Gel electrophoresis and subsequent imaging using phosphorimagers is one of the most important and widely used techniques in RNA and DNA analysis. Radiolabeling nucleic acids with (32)P and detecting bands using a phoshorimager are useful both in a qualitative sense for nucleic acid detection and in a quantitative sense for structural, kinetic, or binding-based assays. Because of this, good resolution of gel bands based on molecular weight and size of RNA or DNA is essential for analysis. The appearance of blurry gel bands of (32)P-labeled RNA and DNA thus represents a serious problem in the laboratory. A quick search on the Internet uncovers numerous reports begrudging the appearance of blurry bands, as well as attempts to fix them without success. Indeed, our laboratories were beset by the intermittent problem of blurry gels for over one year before we found a solution. Herein we describe a simple and cost-effective solution to a problem that we show originates from the phosphorimager cassettes rather than the integrity of the gel itself. We hope that the information provided here will lead to immediate help for other laboratories experiencing similar issues with labeled nucleic acid gel-based assays. The improvement in the clarity of the gels is nothing short of astonishing in many instances and will lead to higher resolution images for analysis and publications.
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spelling pubmed-51132122017-12-01 Eliminating blurry bands in gels with a simple cost-effective repair to the gel cassette Bingaman, Jamie L. Frankel, Erica A. Hull, Chelsea M. Leamy, Kathleen A. Messina, Kyle J. Mitchell, David Park, Hongmarn Ritchey, Laura E. Babitzke, Paul Bevilacqua, Philip C. RNA Methods Note Gel electrophoresis and subsequent imaging using phosphorimagers is one of the most important and widely used techniques in RNA and DNA analysis. Radiolabeling nucleic acids with (32)P and detecting bands using a phoshorimager are useful both in a qualitative sense for nucleic acid detection and in a quantitative sense for structural, kinetic, or binding-based assays. Because of this, good resolution of gel bands based on molecular weight and size of RNA or DNA is essential for analysis. The appearance of blurry gel bands of (32)P-labeled RNA and DNA thus represents a serious problem in the laboratory. A quick search on the Internet uncovers numerous reports begrudging the appearance of blurry bands, as well as attempts to fix them without success. Indeed, our laboratories were beset by the intermittent problem of blurry gels for over one year before we found a solution. Herein we describe a simple and cost-effective solution to a problem that we show originates from the phosphorimager cassettes rather than the integrity of the gel itself. We hope that the information provided here will lead to immediate help for other laboratories experiencing similar issues with labeled nucleic acid gel-based assays. The improvement in the clarity of the gels is nothing short of astonishing in many instances and will lead to higher resolution images for analysis and publications. Cold Spring Harbor Laboratory Press 2016-12 /pmc/articles/PMC5113212/ /pubmed/27852929 http://dx.doi.org/10.1261/rna.059303.116 Text en © 2016 Bingaman et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society http://creativecommons.org/licenses/by-nc/4.0/ This article is distributed exclusively by the RNA Society for the first 12 months after the full-issue publication date (see http://rnajournal.cshlp.org/site/misc/terms.xhtml). After 12 months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.
spellingShingle Methods Note
Bingaman, Jamie L.
Frankel, Erica A.
Hull, Chelsea M.
Leamy, Kathleen A.
Messina, Kyle J.
Mitchell, David
Park, Hongmarn
Ritchey, Laura E.
Babitzke, Paul
Bevilacqua, Philip C.
Eliminating blurry bands in gels with a simple cost-effective repair to the gel cassette
title Eliminating blurry bands in gels with a simple cost-effective repair to the gel cassette
title_full Eliminating blurry bands in gels with a simple cost-effective repair to the gel cassette
title_fullStr Eliminating blurry bands in gels with a simple cost-effective repair to the gel cassette
title_full_unstemmed Eliminating blurry bands in gels with a simple cost-effective repair to the gel cassette
title_short Eliminating blurry bands in gels with a simple cost-effective repair to the gel cassette
title_sort eliminating blurry bands in gels with a simple cost-effective repair to the gel cassette
topic Methods Note
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5113212/
https://www.ncbi.nlm.nih.gov/pubmed/27852929
http://dx.doi.org/10.1261/rna.059303.116
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