title: Eliminating blurry bands in gels with a simple cost-effective repair to the gel cassette.
abstract: 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 with32P 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 of32P-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.
authors.affiliation: Department of Chemistry, The Pennsylvania State University, University Park, Pennsylvania 16802, USA.|Center for RNA Molecular Biology, The Pennsylvania State University, University Park, Pennsylvania 16802, USA.
authors.forename: Jamie L
authors.initials: JL
authors.lastname: Bingaman
authors.affiliation: Department of Chemistry, The Pennsylvania State University, University Park, Pennsylvania 16802, USA.|Center for RNA Molecular Biology, The Pennsylvania State University, University Park, Pennsylvania 16802, USA.
authors.forename: Erica A
authors.initials: EA
authors.lastname: Frankel
authors.affiliation: Department of Chemistry, The Pennsylvania State University, University Park, Pennsylvania 16802, USA.|Center for RNA Molecular Biology, The Pennsylvania State University, University Park, Pennsylvania 16802, USA.
authors.forename: Chelsea M
authors.initials: CM
authors.lastname: Hull
authors.affiliation: Department of Chemistry, The Pennsylvania State University, University Park, Pennsylvania 16802, USA.|Center for RNA Molecular Biology, The Pennsylvania State University, University Park, Pennsylvania 16802, USA.
authors.forename: Kathleen A
authors.initials: KA
authors.lastname: Leamy
authors.affiliation: Department of Chemistry, The Pennsylvania State University, University Park, Pennsylvania 16802, USA.|Center for RNA Molecular Biology, The Pennsylvania State University, University Park, Pennsylvania 16802, USA.
authors.forename: Kyle J
authors.initials: KJ
authors.lastname: Messina
authors.affiliation: Department of Chemistry, The Pennsylvania State University, University Park, Pennsylvania 16802, USA.|Center for RNA Molecular Biology, The Pennsylvania State University, University Park, Pennsylvania 16802, USA.
authors.forename: David
authors.initials: D
authors.lastname: Mitchell
authors.affiliation: Center for RNA Molecular Biology, The Pennsylvania State University, University Park, Pennsylvania 16802, USA.|Department of Biochemistry and Molecular Biology, Pennsylvania State University, University Park, Pennsylvania 16802, USA.
authors.forename: Hongmarn
authors.initials: H
authors.lastname: Park
authors.affiliation: Department of Chemistry, The Pennsylvania State University, University Park, Pennsylvania 16802, USA.|Center for RNA Molecular Biology, The Pennsylvania State University, University Park, Pennsylvania 16802, USA.
authors.forename: Laura E
authors.initials: LE
authors.lastname: Ritchey
authors.affiliation: Center for RNA Molecular Biology, The Pennsylvania State University, University Park, Pennsylvania 16802, USA.|Department of Biochemistry and Molecular Biology, Pennsylvania State University, University Park, Pennsylvania 16802, USA.
authors.forename: Paul
authors.initials: P
authors.lastname: Babitzke
authors.affiliation: Department of Chemistry, The Pennsylvania State University, University Park, Pennsylvania 16802, USA.|Center for RNA Molecular Biology, The Pennsylvania State University, University Park, Pennsylvania 16802, USA.|Department of Biochemistry and Molecular Biology, Pennsylvania State University, University Park, Pennsylvania 16802, USA.
authors.forename: Philip C
authors.initials: PC
authors.lastname: Bevilacqua
mesh_terms: D003362:Cost-Benefit Analysis; D004247:DNA / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D012313:RNA / Q000737:chemistry*
chemical_list: D012313:RNA; D004247:DNA
keywords: PAGE; gel electrophoresis; methods