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

Author:
Beckmann, Benedikt M., Grünweller, Arnold, Weber, Michael H.W., Hartmann, Roland K.
Source:
Nucleic acids research 2010 v.38 no.14 pp. e147
ISSN:
0305-1048
Subject:
Northern blotting, RNA, bacteria, crosslinking, polyacrylamide gel electrophoresis
Abstract:
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.
Agid:
2272349