BIL 250 Lecture Notes - Lecture 5: Electrophoretic Mobility Shift Assay, Human Genome Project, Dna Sequencing
18 Apr 2017
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Dna sequencing is the ultimate way to characterize dna structure at the molecular level. Dideoxynucleotide: deoxynucleotide with a hydrogen at 3 instead of an oh. Dideoxynucleotide causes dna synthesis to terminate when get randomly incorporated into the polynucleotide chain. Since early 1990s, dna sequencing has been done through computer automated sanger reaction based technology. This procedure can determine if a protein or mixture of proteins is capable of binding to a given dna or rna sequence. Study of dna protein interactions useful for understanding transcription factor activity and histone placement. Also called gel mobility shift assay or electrophoretic mobility shift assay. Binding of a protein to a fragment of dna retards its rate of movement through a gel. Must be performed under non-denaturing conditions as unfolding of protein or separation of dna strands would cause it to release dna protein interaction. Complexes that migrate more slowly indicate that dna was bound by protein.
