LAB5_OCON - In the plasmid DNA isolation procedure, the...

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Caroline O’Connor Lab 4 T.A. James Peterman TITLE: Mitochondrial DNA PCR Lab 1.) Cheek cells were used because they are amongst the easiest cells in our body to isolate, and can be easily scraped or swished off and collected. Because they are cells from human DNA, they are eukaryotic and have a nucleus and mitochondrion. Therefore, we can isolate the DNA and induce a PCR reaction. 2.) If you wanted to study variations in a gene that was normally expressed in muscle cells, you could still use DNA from cheek cells. This is because all of the cells in the body, with the exception of the sperm and egg cells) have the same DNA. Therefore, we can isolate mitochondrial DNA from any cell. 3.) The mitochondrial DNA isolation procedure differed from the plasmid DNA isolation procedure in the following ways: In the plasmid DNA isolation procedure, it was bacterial plasmid DNA that was isolated whereas in the mitochondrial DNA isolation procedure, the DNA was not a derivative of a bacteria
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Unformatted text preview: In the plasmid DNA isolation procedure, the cell wall was broken down by lysozyme and burst open by heat, whereas in the mitochondrial DNA procedure, the cell wall was burst open by using heat alone • In the plasmid DNA isolation procedure, plasmid DNA was grown in large amounts through transformation of a bacteria that will divide and the DNA will be copied with every division. In the mitochondrial DNA isolation procedure, a polymerase chain reaction was used to amplify the mitochondrial control region and thus produce large amounts of mitochondrial DNA The procedures were similar in the following ways: • A centrifuge or microfuge were used in both for DNA isolation • Heat was used in both to aid in the rupturing of the cell wall • Mitochondria contain a circular piece of DNA similar to a plasmid containing some but not all of the genes for producing mitochondrial RNAs and, as a result, proteins • Mitochondrial DNA uses similar codon usage as bacteria...
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This note was uploaded on 08/11/2010 for the course IPHY 3060 taught by Professor Allen,davi during the Fall '09 term at Colorado.

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