Practice question

EMSA is excellent for qualitative and semi-quantitative evaluation of sequence-specific DNA-protein complex formation using short labeled probes of known sequence. Under non-denaturing conditions, binding affinity, specificity via competitor DNA and complex stoichiometry can be assessed. However, assay fundamentally depends on prior knowledge of DNA fragment sequence and cannot determine unknown nucleotide order. Determination of primary structure requires Sanger sequencing or next-generation sequencing technologies. Consequently, EMSA is unsuitable for DNA sequence determination, although it remains invaluable for visualizing protein-DNA complexes and measuring binding affinity.