BIOL 4374 Lecture Notes - Lecture 10: Dna Profiling, Intron, Hydrolysis
22 Feb 2017
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Lecture 10/11: can create new dna sequences in vitro that could be tested in vivo. Hello molecular biology: before this, selective breeding was the only way to control dna, made possible via discovery: restriction nuclease. Bacterial enzymes that cut foreign dna at specific elements. Bacterial dna is safe because specific dna sequences are chemically modified. Dna is cut by hydrolysis of phosphodiester bonds. Blunt ends: even cut thru the double helix. Sticky ends: uneven cut thru the double helix. Can be to blunt ends: gel electrophoresis: separate dna fragment by size. Agarose is commonly used because it is cheap. Detect dna via: fluorescent dye, 32p labeling. Radioactive phosphorus labeling is the most sensitive. A piece of dna can be isolated as it migrates through the gel: cut dna- linear, has a predictable size, uncut dna. Nicked/relaxed circle: slowest on agarose gel: detect specific dna sequences through hybridization probe. The probe is generally dna stable.
