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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978-1-4939-5941-9Springer Science+Business Media, LLC 2012

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Bioinformatic Discovery of Bacterial Regulatory RNAs Using SIPHT searching for regions of intergenic sequence conservation upstream of predicted intrinsic transcription terminators. Each locus is then annotated for numerous features that provide clues about its potential function and/or enable the most reliable candidates to be identified.

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Crystallization of RNA–Protein Complexes: From Synthesis and Purification of Individual Components tin vitro, and proteins that were overexpressed in . and purified to be RNase-free. The complex was crystallized using a sitting drop setup; initial screening for suitable crystallization conditions was performed using a sparse matrix approach.

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Anlagen zur Start- und Zündhilfe 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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Anlagen zur Start- und Zündhilfeg in terms of stoichiometry, affinity, and heat (enthalpy), while DSC can provide RNA stability in terms of heat capacity, melting temperature, and folding enthalpy. Here, we offer detailed experimental protocols for studying such RNA systems with commercially available conventional and high-throughput ITC and DSC instruments.

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https://doi.org/10.1007/978-3-662-07709-2ative 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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Geschichte und Grundlagen des Dieselmotorsucleotides, 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.