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Titlebook: Bacterial Regulatory RNA; Methods and Protocol Kenneth C. Keiler Book 2012 Springer Science+Business Media, LLC 2012 RNA.RNA polymerase.bac

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RNA Visualization in Bacteria by Fluorescence In Situ Hybridization microscopy. This chapter describes the use of FISH to visualize tmRNA, a regulatory RNA required for .-translation. The method can be adapted to visualize the localization of other regulatory and messenger RNAs as well.
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Gel Mobility Shift Assays to Detect Protein–RNA Interactionsative polyacrylamide gels in Tris/borate/EDTA buffer, although an alternative Tris-glycine buffering system is superior in many situations. Here, we describe both gel shift methods, along with strategies to improve separation of protein–RNA complexes from free RNA, which can be a particular challenge for small RNA binding proteins.
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RNase Footprinting of Protein Binding Sites on an mRNA Target of Small RNAsucleotides, to footprint the protein Hfq on the . mRNA leader. This protocol covers how to form the RNP complex, determine the correct dose of enzyme, footprint the protein, and analyze the cleavage pattern using primer extension.
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Use of Semi-quantitative Northern Blot Analysis to Determine Relative Quantities of Bacterial CRISPR stabilities of both pre-crRNA and crRNA. The procedures described in this chapter can be used with very minor modifications to monitor the abundance and stabilities of transcripts of various lengths from many bacterial sources.
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