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BIO 302 Genetics

DNA A.G.E.

QuestionAnswer
Agarose is a ------? Protein, polysaccharide, nucleic acid, or lipid bi-layer? Polysacchradide
What is the charge density of a molecule? Charge per mass
Agarose electrophoresis of DNA molecules separates molecules based on -----? size
In electrophoresis, What is a band? Group of molecules in a gel migrating at the same time and rate.
What is the purpose of the following in the DNA sample buffer: glycerol, TRIS, EDTA (2 parts) Glycerol: weighs down sample. Water soluble and more dense than water TRIS:Weak Base (pH Buffer) EDTA: Weak Acid (pH Buffer) Chelates divalent cations required by DNAses. ( meaning that they bind Ca+, Mg+, Mn+)
What is a tracking dye? IS NOT THE STAIN>>> negatively charged molecule that moves at known base pair equivalent rate.
DNA molecules will move toward the positive or negative electrode during electrophoresis. Why? positive, because moves toward opposite charge and DNA bc of phosphorus has overall negative charge.
Describe an advantage of to using high voltage electrophoresis. Speed
Describe a disadvantage of using high voltage electrophoresis. Overheats and breaks down matrix bc of resistance causing band fuzziness. This decreases resolution.
What is band resolution? The ability to distinguish two bands as separate.
To separate small DNA molecules it is best to use a high or low concentration of agrarose? High
Ethidium Bromide intercalates DNA and is a fluorescent dye. Explain terms. Intercalates: Ability to slide in between the nucleotides of DNA bc it is flat. Fluorescence: Converts the energy to light energy. Produces light this way.
DNA agarose gel electrophoresis may be employed to determine the size of an unknown fragment. What are two other ways to use DNA AGE? 1) DNA Fingerprinting 2) Purify a particular DNA (separate it out)
Created by: michellerush630
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