Electrophoretic Mobility Shift Assay of DNA and CRISPR-Cas Ribonucleoprotein Complexes

T.A. Künne, E.R. Westra, S.J.J. Brouns

Research output: Chapter in Book/Report/Conference proceedingChapterAcademicpeer-review

6 Citations (Scopus)

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
Original languageEnglish
Title of host publicationCRISPR : Methods and Protocols
EditorsM. Lundgren, E. Charpentier, P.C. Fineran
Pages171-184
Number of pages371
DOIs
Publication statusPublished - 2015

Publication series

NameMethods Molecular Biology
PublisherSpringer Humana Press
Number1311

Keywords

  • Affi nity
  • Binding assay
  • Cascade
  • CRISPR
  • Electrophoretic mobility shift assay
  • EMSA
  • Gel shift
  • Protein–DNA interaction
  • RNA guide

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