530L_notes_lecture2

530L_notes_lecture2 - Lecture 2 III Agarose gel...

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Lecture 2 III. Agarose gel electrophoresis: a method for visualizing nucleic acids Method for visualizing DNA into lesser extent RNA Common technique A. What is agarose? Carbohydrate derived from algae Repeating disaccharide units – long polysaccharide B. Physical Properties of Agarose (see ppt slide) Insoluble “white powder” Have to heat agarose in solution Heat causes agarose to physically change conformation into randomly coiled structure Ex. Jello After cooling, the randomly coiled structure changes link up and interact with one another, double helical structure with a pore C. Solid agarose- a mesh-like matrix of PORES (see ppt slide). These pores are: 1. large enough for nucleic acids to be pushed through 2. defined size 3. depending on how much agarose into buffer when making the gel, can control the size of the pores To make agarose gel: Weight powder agarose per volume D. Factors that influence the migration of nucleic acids through a gel during electrophoresis All n.a. have the same charge at certain ph
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530L_notes_lecture2 - Lecture 2 III Agarose gel...

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