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Ultra-fast conductive media for RNA electrophoretic mobility shift assays

The use of RNA electrophoretic mobility shift assays (REMSAs) for analysis of RNA–protein interactions have been limited to lengthy assay time and qualitative assessment. To vastly improve assay efficiency, feasibility and quality of data procured from REMSAs, we combine here some of the best-known...

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Autores principales: Brown, Samantha Z, Agostini, Lebaron C, Thomsett, Henry L, Brody, Jonathan R
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Future Science Ltd 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7026733/
https://www.ncbi.nlm.nih.gov/pubmed/31870164
http://dx.doi.org/10.2144/btn-2019-0111
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author Brown, Samantha Z
Agostini, Lebaron C
Thomsett, Henry L
Brody, Jonathan R
author_facet Brown, Samantha Z
Agostini, Lebaron C
Thomsett, Henry L
Brody, Jonathan R
author_sort Brown, Samantha Z
collection PubMed
description The use of RNA electrophoretic mobility shift assays (REMSAs) for analysis of RNA–protein interactions have been limited to lengthy assay time and qualitative assessment. To vastly improve assay efficiency, feasibility and quality of data procured from REMSAs, we combine here some of the best-known labeling and electrophoretic techniques. Nucleic acid fragments are end-labeled with fluorescent tags, as opposed to the radioactive or biotin tags. The fluorescent probes may be detected directly from the electrophoresis gel, eliminating the need for cumbersome membrane transfer and immunoblotting. Modifying the REMSA protocol to include low-molarity, lithium borate conductive media and near-infrared-labeled probes allows for a reduction assay time, quantitative comparison between experimental conditions and crisp band resolution (i.e., optimized results).
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spelling pubmed-70267332020-02-27 Ultra-fast conductive media for RNA electrophoretic mobility shift assays Brown, Samantha Z Agostini, Lebaron C Thomsett, Henry L Brody, Jonathan R Biotechniques Benchmark The use of RNA electrophoretic mobility shift assays (REMSAs) for analysis of RNA–protein interactions have been limited to lengthy assay time and qualitative assessment. To vastly improve assay efficiency, feasibility and quality of data procured from REMSAs, we combine here some of the best-known labeling and electrophoretic techniques. Nucleic acid fragments are end-labeled with fluorescent tags, as opposed to the radioactive or biotin tags. The fluorescent probes may be detected directly from the electrophoresis gel, eliminating the need for cumbersome membrane transfer and immunoblotting. Modifying the REMSA protocol to include low-molarity, lithium borate conductive media and near-infrared-labeled probes allows for a reduction assay time, quantitative comparison between experimental conditions and crisp band resolution (i.e., optimized results). Future Science Ltd 2019-12-24 2019-12 /pmc/articles/PMC7026733/ /pubmed/31870164 http://dx.doi.org/10.2144/btn-2019-0111 Text en © 2019 Jonathan R. Brody This work is licensed under the Attribution-NonCommercial-NoDerivatives 4.0 Unported License (http://creativecommons.org/licenses/by-nc-nd/4.0/)
spellingShingle Benchmark
Brown, Samantha Z
Agostini, Lebaron C
Thomsett, Henry L
Brody, Jonathan R
Ultra-fast conductive media for RNA electrophoretic mobility shift assays
title Ultra-fast conductive media for RNA electrophoretic mobility shift assays
title_full Ultra-fast conductive media for RNA electrophoretic mobility shift assays
title_fullStr Ultra-fast conductive media for RNA electrophoretic mobility shift assays
title_full_unstemmed Ultra-fast conductive media for RNA electrophoretic mobility shift assays
title_short Ultra-fast conductive media for RNA electrophoretic mobility shift assays
title_sort ultra-fast conductive media for rna electrophoretic mobility shift assays
topic Benchmark
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7026733/
https://www.ncbi.nlm.nih.gov/pubmed/31870164
http://dx.doi.org/10.2144/btn-2019-0111
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