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DNA Profiling Lab

by: Shanell Coleman

DNA Profiling Lab 18519

Shanell Coleman
GPA 3.7

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Lab # 4 DNA Profiling Lab
Biology 1101
Thomas Buxton
Class Notes
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This 2 page Class Notes was uploaded by Shanell Coleman on Monday October 10, 2016. The Class Notes belongs to 18519 at Augusta State University taught by Thomas Buxton in Fall 2016. Since its upload, it has received 7 views. For similar materials see Biology 1101 in Science at Augusta State University.


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Date Created: 10/10/16
Lab #4  DNA Profiling Lab  1. Supernatant: denoting the liquid lying above a solid residue after  crystallization, precipitation, centrifugation, or other process 2. Soap: destroying all cell membranes, allowing the cell contents to spill out  (cell lysis). 3. DNA is only soluble at a pH near physiological levels. The baking soda  serves as a buffering system that raises the pH and releases the DNA from  bound proteins.  4. The meat tenderizer: is an enzyme (proteinase) that removes proteins  bound to the DNA and destroys enzymes (endonucleases) which would  chew up the DNA.  5.  Since the supernatant is thick from the cellular contents, carefully pouring  the alcohol on top of the supernatant leaves two distinct layers. DNA is  soluble in water, but not in alcohol; thus, the DNA present at the water –  alcohol interface precipitates out of solution, allowing it to be seen.  Gel Electrophoresis Gel electrophoresis is a basic biotechnology technique that separates  macromolecules according to their size and charge. It is frequently used to  analyze a manipulate samples of DNA, RNA, or proteins. In gel electrophoresis, samples to be separated are applied to a porous gel medium made of a material  such as agarose. Agarose is a purified form of agar, a gelatinous substance  extracted from red algae.  All the samples are loaded into wells of the agarose gel. The gel is then placed in a  chamber that is connected to a power supply. To “run” the gel an electrical  current is applied to the gel. The chamber is designed with a positive electrode  (anode) at one end and a negative electrode (cathode) at the other end.  Electrophoresis literally means “to carry with electricity,” once the electric  field is established, charged molecules in the samples migrate through the  pores of the gel toward their pole of attraction. Molecules with a net negative  charge migrate toward the positive electrode and molecules with a net positive  charge migrate toward the negative electrode. The overall charge of a molecule  affects the speed at which it travels through the gel. Highly charged molecules  migrate more quickly through the gel than weakly charged molecules.  The mobility of a molecule during gel electrophoresis also depends on its  molecular size and shape. The small pores of the gel matrix act as a sieve that  provides great resolving power. Small molecules maneuver more easily through  the pores than larger molecules and therefore travel relatively quickly. Large  molecules encounter more resistance as they make their way through the tiny pores and therefore travel at a slower rate.  In order to see the DNA in the gel, a chemical called, ethidium bromide (EtBr) is added to the agarose while it was still in its molten state. Ethidium bromide is a molecule that inserts or intercalates between the bases of double stranded DNA.  Under UV light EtBr fluoresces, therefore DNA becomes visible under UV light in the presence of EtBr. EtBr is a potent mutagen (may cause genetic damage), and is  moderately toxic.  Once the gel has run and a picture of the DNA bands has been taken, you can  analyze the size of the DNA bands by comparing it to the lay containing the DNA  ladder. A DNA ladder is a solution that contains known sizes of SNA. You  compare your samples with the known bands to estimate the size of your DNA  bands.  A  T = Adenine  Thymine  C  G = Cytosine  Guanine  Bind by hydrogen bonds: each is coiled from double helix nucleotides also are the  monomers of nucleic acids.  Nucleotides have three components:  1. Phosphate group (same for DNA and RNA) 2. Pentose (5­carbons, different sugar for DNA and RNA)  3. Nitrogen – containing base (defines which nucleotide)


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