01:447:380 Lecture Notes - Lecture 19: Agarose Gel Electrophoresis, Southern Blot, Dystrophin
Document Summary
General methods: restriction enzyme digest, each enzyme recognizes a specific dna sequence a) Using gel electrophoresis: dna moves from (-) to (+, smaller dna pieces more faster, result: gel with many different sized dna fragments, gel electrophoresis. Probe binding confirms that sequence of interest tag/radioactivity) is in sample that was loaded on gel: signals on filter paper corresponds to size of dna fragments, can be used to detect homologous genes in different species. Example: dystrophy in humans. you want to determine if gorillas, chimpanzees, and bonobos have a gene homologous to the human dystrophin gene. Southern blot using dna from all species. Purify dna from cells of gorilla, chimp, and bonobo. Cut dna with restriction endonuclease creates smaller dna fragments. Run agarose gel; transfer dna to filter paper separated by size. Probe blot using ssdna based on conserved portion of human dystrophin gene use conditions where probe can bind to sequence that may not be exact complement.
