BTEC 4P11 Lecture Notes - Sybr Green I, Wild Type, Geforce 3 Series
Document Summary
The interaction of proteins with dna is central to the control of many cellular processes including dna replication, recombination and repair, transcription, and viral assembly. One technique that is central to studying gene regulation and determining protein:dna interactions is the electrophoretic mobility shift assay (emsa). The emsa technique is based on the observation that protein:dna complexes migrate more slowly than free dna molecules when subjected to non-denaturing polyacrylamide or agarose gel electrophoresis. Because the rate of dna migration is shifted or retarded upon protein binding, the assay is also referred to as a gel shift or gel retardation assay. An advantage of studying dna:protein interactions by an electrophoretic assay is the ability to resolve complexes of different stoichiometry or conformation. Another major advantage for many applications is that the source of the dna-binding protein may be a crude nuclear or whole cell extract rather than a purified preparation.