By Jonathan B. Chaires

This quantity consolidates the most important tools for learning ligand-nucleic acid interactions right into a handy resource. thoughts which are tested variety from biophysical and chemical methods to tools rooted in molecular and phone biology.

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Bajic, D. W. Boykin, and W. D. Wilson, Proc. Natl. Acad. Sci. A. 97, 12 (2000). -O. Markgren, M. Hamalained, and U. H. Danielson,Anal. Biochem. 265, 340 (1998). "Biacore, Uppsala, Sweden, June 1994. [2] RNA INTERACTIONS Description of Surface Plasmon Resonance 27 Biosensors SPR biosensors employ surface plasmon resonance to qualitatively and/or quantitatively describe molecular interactions. Although several companies now offer SPR biosensors, 54 currently, instruments from BIAcore (Uppsala, Sweden) are the most widely used.

Some researchers block the remaining biotin binding sites and the biotin binding sites on the streptavidin in the control flow cell with free biotin. This will depend on any nonspecific interactions between components in the flow solution with streptavidin on the sensor chip surface. STORAGEOF RNA SENSOR CHIP. Researchers at the NIH store oligonucleotide sensor chips in Tris-buffered saline (TBS) buffer to maintain binding activity (Inna Gorshkova, personal communication, 2000). To do so, remove the sensor chip from the white cartridge, being careful not to touch the flow cells.

1. Dock the streptavidin-coated sensor chip into the instrument. 2. Prime the instrument three times with HBS buffer (note that priming does not need to be done if the streptavidin chip was just made as outlined above, 36 BIOPHYSICALAPPROACHES [2] has not been removed from the instrument and HBS has been the running buffer). 3. Start a sensorgram with a 20-/zl/min flow rate. 4. Inject activation buffer for 1 min (20 ~1) three times to remove any unbound streptavidin from the sensor chip. 5. Allow buffer to flow for at least 5 min before immobilizing the RNA.

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