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Electrophoretic Mobility Shift Assay of DNA and CRISPR-Cas Ribonucleoprotein Complexes.
Künne, Tim; Westra, Edze R; Brouns, Stan J J.
Afiliación
  • Künne T; Laboratory of Microbiology, Department of Agrotechnology and Food Sciences, Wageningen University, Dreijenplein 10, 6703 HB, Wageningen, The Netherlands.
Methods Mol Biol ; 1311: 171-84, 2015.
Article en En | MEDLINE | ID: mdl-25981473
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.
Asunto(s)

Texto completo: 1 Banco de datos: MEDLINE Asunto principal: Ribonucleoproteínas / ADN / Proteínas de Escherichia coli / Ensayo de Cambio de Movilidad Electroforética / Sistemas CRISPR-Cas Idioma: En Revista: Methods Mol Biol Asunto de la revista: BIOLOGIA MOLECULAR Año: 2015 Tipo del documento: Article País de afiliación: Países Bajos

Texto completo: 1 Banco de datos: MEDLINE Asunto principal: Ribonucleoproteínas / ADN / Proteínas de Escherichia coli / Ensayo de Cambio de Movilidad Electroforética / Sistemas CRISPR-Cas Idioma: En Revista: Methods Mol Biol Asunto de la revista: BIOLOGIA MOLECULAR Año: 2015 Tipo del documento: Article País de afiliación: Países Bajos