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

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′-digoxigeninendlabeled for non-radioactive probe detection with 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimidemediated chemical crosslinking of RNAs to nylon membranes, and necessitates the use of native PAGE either with the TBE or MOPS buffer system. © The Author(s) 2010. Published by Oxford University Press.