Biophysical, Chemical, and Functional Probes of RNA by Daniel Herschlag

By Daniel Herschlag

This MIE quantity offers laboratory recommendations that objective to foretell the constitution of a protein that could have large implications starting from drug layout, to mobile pathways and their dynamics, to viral access into cells.
Expert researchers introduce the main complicated applied sciences and strategies in protein constitution and folding
Includes strategies on tiling assays.

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Additional info for Biophysical, Chemical, and Functional Probes of RNA Structure, Interactions and Folding: Part B

Example text

Transcription from this DNA results in the desired RNA product linked to the affinity purification tag. At right is an ethidium bromide stained agarose gel showing representative results from reactions 1a, 1b, and 2. Reprinted with permission from Edwards et al. (2009). 10 Amanda Y. Keel et al. 2. Procedures 1. PCR reaction 1a (50 ml): This reaction generates a DNA product that contains the sequence of the desired RNA under control of the T7 promoter, as well as a ‘‘joint’’ on the 30 -end that allows it to be combined with the product of reaction 1b (in reaction 2) to generate the full-length product.

1987). Oligoribonucleotide synthesis using T7 RNA polymerase and synthetic DNA templates. Nucleic Acids Res. 15, 8783–8798. Pestova, T. , Shatsky, I. , Fletcher, S. , Jackson, R. , and Hellen, C. U. (1998). A prokaryotic-like mode of cytoplasmic eukaryotic ribosome binding to the initiation codon during internal translation initiation of hepatitis C and classical swine fever virus RNAs. Genes Dev. 12, 67–83. , and Breaker, R. R. (2006). Characteristics of the glmS ribozyme suggest only structural roles for divalent metal ions.

Jansen, J. , Soukup, J. , and Soukup, G. A. (2005). Ligand requirements for glmS ribozyme self-cleavage. Chem. Biol. 12, 1221–1226. Native Purification of RNAs 25 McKenna, S. , Puglisi, E. , Lindhout, D. , Aitken, C. , Marshall, R. , and Puglisi, J. D. (2007). Purification and characterization of transcribed RNAs using gel filtration chromatography. Nat. Protoc. 2, 3270–3277. Milligan, J. , Groebe, D. , Witherell, G. , and Uhlenbeck, O. C. (1987). Oligoribonucleotide synthesis using T7 RNA polymerase and synthetic DNA templates.

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