Northern blot detection of endogenous small RNAs (∼14 nt) in bacterial total RNA extracts

Here we describe a northern blot procedure that allows the detection of endogenous RNAs as small as ∼14 nt in total RNA extracts from bacteria. RNAs that small and as part of total bacterial RNA extracts usually escape detection by northern blotting. The approach combines LNA probes 5′-digoxigenin-e...

Descripción completa

Detalles Bibliográficos
Autores principales: Beckmann, Benedikt M., Grünweller, Arnold, Weber, Michael H. W., Hartmann, Roland K.
Formato: Texto
Lenguaje:English
Publicado: Oxford University Press 2010
Materias:
Methods Online
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2919735/
https://www.ncbi.nlm.nih.gov/pubmed/20504856
http://dx.doi.org/10.1093/nar/gkq437
_version_ 1782185218131623936
author Beckmann, Benedikt M.
Grünweller, Arnold
Weber, Michael H. W.
Hartmann, Roland K.
author_facet Beckmann, Benedikt M.
Grünweller, Arnold
Weber, Michael H. W.
Hartmann, Roland K.
author_sort Beckmann, Benedikt M.
collection PubMed
description Here we describe a northern blot procedure that allows the detection of endogenous RNAs as small as ∼14 nt in total RNA extracts from bacteria. RNAs that small and as part of total bacterial RNA extracts usually escape detection by northern blotting. The approach combines LNA probes 5′-digoxigenin-endlabeled for non-radioactive probe detection with 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide-mediated chemical crosslinking of RNAs to nylon membranes, and necessitates the use of native PAGE either with the TBE or MOPS buffer system.
format Text
id pubmed-2919735
institution National Center for Biotechnology Information
language English
publishDate 2010
publisher Oxford University Press
record_format MEDLINE/PubMed
spelling pubmed-29197352010-08-11 Northern blot detection of endogenous small RNAs (∼14 nt) in bacterial total RNA extracts Beckmann, Benedikt M. Grünweller, Arnold Weber, Michael H. W. Hartmann, Roland K. Nucleic Acids Res Methods Online Here we describe a northern blot procedure that allows the detection of endogenous RNAs as small as ∼14 nt in total RNA extracts from bacteria. RNAs that small and as part of total bacterial RNA extracts usually escape detection by northern blotting. The approach combines LNA probes 5′-digoxigenin-endlabeled for non-radioactive probe detection with 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide-mediated chemical crosslinking of RNAs to nylon membranes, and necessitates the use of native PAGE either with the TBE or MOPS buffer system. Oxford University Press 2010-08 2010-05-26 /pmc/articles/PMC2919735/ /pubmed/20504856 http://dx.doi.org/10.1093/nar/gkq437 Text en © The Author(s) 2010. Published by Oxford University Press. http://creativecommons.org/licenses/by-nc/2.5 This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Methods Online
Beckmann, Benedikt M.
Grünweller, Arnold
Weber, Michael H. W.
Hartmann, Roland K.
Northern blot detection of endogenous small RNAs (∼14 nt) in bacterial total RNA extracts
title Northern blot detection of endogenous small RNAs (∼14 nt) in bacterial total RNA extracts
title_full Northern blot detection of endogenous small RNAs (∼14 nt) in bacterial total RNA extracts
title_fullStr Northern blot detection of endogenous small RNAs (∼14 nt) in bacterial total RNA extracts
title_full_unstemmed Northern blot detection of endogenous small RNAs (∼14 nt) in bacterial total RNA extracts
title_short Northern blot detection of endogenous small RNAs (∼14 nt) in bacterial total RNA extracts
title_sort northern blot detection of endogenous small rnas (∼14 nt) in bacterial total rna extracts
topic Methods Online
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2919735/
https://www.ncbi.nlm.nih.gov/pubmed/20504856
http://dx.doi.org/10.1093/nar/gkq437
work_keys_str_mv AT beckmannbenediktm northernblotdetectionofendogenoussmallrnas14ntinbacterialtotalrnaextracts
AT grunwellerarnold northernblotdetectionofendogenoussmallrnas14ntinbacterialtotalrnaextracts
AT webermichaelhw northernblotdetectionofendogenoussmallrnas14ntinbacterialtotalrnaextracts
AT hartmannrolandk northernblotdetectionofendogenoussmallrnas14ntinbacterialtotalrnaextracts

Ejemplares similares