Northern blot detection of endogenous small RNAs (~14 nt) in bacterial total RNA extracts |
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Authors: | Benedikt M Beckmann Arnold Grünweller Michael H W Weber Roland K Hartmann |
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Institution: | Institut für Pharmazeutische Chemie, Philipps-Universität Marburg, D-35037 Marburg, Germany |
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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. |
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