title: Electrophoretic Mobility Shift Assay of DNA and CRISPR-Cas Ribonucleoprotein Complexes.
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 (Kd). Here, we describe how two types of EMSA assays are performed using the Cascade ribonucleoprotein complex from Escherichia coli as an example.
authors.affiliation: Laboratory of Microbiology, Department of Agrotechnology and Food Sciences, Wageningen University, Dreijenplein 10, 6703 HB, Wageningen, The Netherlands.
authors.forename: Tim
authors.initials: T
authors.lastname: Künne
authors.forename: Edze R
authors.initials: ER
authors.lastname: Westra
authors.forename: Stan J J
authors.initials: SJ
authors.lastname: Brouns
mesh_terms: D064113:CRISPR-Cas Systems; D004247:DNA / Q000378:metabolism*; D024202:Electrophoretic Mobility Shift Assay / Q000379:methods*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D007091:Image Processing, Computer-Assisted; D009841:Oligonucleotides / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D012261:Ribonucleoproteins / Q000235:genetics / Q000378:metabolism*; D013194:Staining and Labeling
chemical_list: D029968:Escherichia coli Proteins; D009841:Oligonucleotides; D012261:Ribonucleoproteins; D004247:DNA