Titlebook: Recombinant and In Vitro RNA Synthesis; Methods and Protocol Graeme L. Conn Book 2012 Springer Science+Business Media, LLC 2012 RNA purific

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Preparative Separation of Ribonucleoside Monophosphates by Ion-Pair Reverse-Phase HPLC,g these, nucleotide-specific and site-specific labeling methods can help tremendously in simplifying complex NMR data, while providing unique opportunities for structural investigation of larger RNAs. Such methods generally require separation of individual isotopically labeled ribonucleoside monopho

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Preparation of Short RNA by In Vitro Transcription,nthesis of very short RNA of 10–20 nucleotides in length by in vitro transcription. These RNA oligomers can be purified conveniently by gel filtration chromatography. We also described how to study RNA and protein binding by gel filtration chromatography.

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Native RNA Purification by Gel Filtration Chromatography,fication using Superdex 75 or 200 gel filtration columns. This approach can be extended to purify biologically interesting RNA complexes such as RNA–protein complexes that have been generated from either synthetic or in vitro transcribed RNAs and recombinant proteins.

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Biosynthetic Preparation of 13C/15N-Labeled rNTPs for High-Resolution NMR Studies of RNAs,hion from bacterial cultures, using common and versatile . strains. This chapter also covers procedures for extraction and digestion of the total RNA from bacterial cells, purification of the ribonucleoside 5′-monophosphates and their enzymatic phosphorylation to rNTPs.

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Splint Ligation of RNA with T4 DNA Ligase,in an RNA and for the assembly of smaller synthetic RNAs into longer RNA molecules. Such modifications enable a wide range of experiments to be carried out with the modified RNA including structural studies, co-immunoprecipitations, and the ability to map sites of RNA:RNA and RNA:protein interactions.

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