Abstract
The Electrophoretic Mobility Shift Assay is a straightforward and inexpensive method for the determination and quantification of protein–nucleic acid interactions. It relies on the different mobility of free and protein-bound nucleic acid in a gel matrix during electrophoresis. Nucleic acid affinities of crRNA-Cas complexes can be quantified by calculating the dissociation constant (K d). Here, we describe how two types of EMSA assays are performed using the Cascade ribonucleoprotein complex from Escherichia coli as an example.
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Acknowledgements
This work was financially supported by a KNAW Beijerinck premium and NWO Vidi grant to S.J.J.B. (864.11.005). ERW received funding from the People Programme (Marie Curie Actions) of the European Union’s Seventh Framework Programme (FP7/2007-2013) under REA grant agreement n0 (327606).
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Künne, T., Westra, E.R., Brouns, S.J.J. (2015). Electrophoretic Mobility Shift Assay of DNA and CRISPR-Cas Ribonucleoprotein Complexes. In: Lundgren, M., Charpentier, E., Fineran, P. (eds) CRISPR. Methods in Molecular Biology, vol 1311. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2687-9_11
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DOI: https://doi.org/10.1007/978-1-4939-2687-9_11
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2686-2
Online ISBN: 978-1-4939-2687-9
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