By Daniel Herschlag
This MIE quantity offers laboratory ideas that objective to foretell the constitution of a protein which could have super 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 methods in protein constitution and folding
Includes options on tiling assays.
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Extra resources for Biophysical, Chemical, and Functional Probes of RNA Structure, Interactions and Folding: Part B
S. (1994). Mutations that increase the affinity of a translational repressor for RNA. Nucleic Acids Res. 22, 3748–3752. Lukavsky, P. , and Puglisi, J. D. (2004). Large-scale preparation and purification of polyacrylamide-free RNA oligonucleotides. RNA 10, 889–893. Lukavsky, P. , Otto, G. , and Puglisi, J. D. (2003). Structure of HCV IRES domain II determined by NMR. Nat. Struct. Biol. 10, 1033–1038. McCarthy, T. , Plog, M. , Floy, S. , Jansen, J. , Soukup, J. , and Soukup, G. A. (2005). Ligand requirements for glmS ribozyme self-cleavage.
12 Amanda Y. Keel et al. 4. 1. 2. Procedures 1. RNA is made by using the PCR-generated DNA template directly in an in vitro transcription. We generally conduct a transcription reaction of 5 ml final volume using the 1 ml PCR reaction 2 DNA from above, using the following conditions: 1Â HEPES transcription buffer 32 mM MgCl2 4 mM each NTP T7 RNA polymerase to a final concentration of $50 mg/ml 1 unit/ml inorganic pyrophosphatase (optional) 1 ml DNA template from PCR reaction 2, above RNase-free water to a final volume of 5 ml 2.
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