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20 Cards in this Set
- Front
- Back
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What is transfection?
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The process of artificially introducing recombinant DNA into eukaryotic cells.
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How is transfection achieved?
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1. DNA is mixed with "transfection reagent"
2. Liposome is incubated with cells and fuses with cells 3. Once the Liposome fuses with the cell membrane, it releases the Transfected DNA into the cells these are normally marked with a florescent protien |
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What serves as the source of recombinant DNA for genomics?
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Plasmid DNA
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How many chromosomes does human DNA have?
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23 pairs
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Who discovered the structure of DNA?
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Watson and Crick
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What is the structure of DNA?
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Double helix composed of nucleotides.
Each nucleotide has three parts: Phospoate group, dioxyribose sugar, and a nucleotide base |
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What are the four nucleotides found in DNA
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Guanine, Adenine, Cytosine, Thaymine
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What are the steps to isolate DNA?
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1. Lyse cells open to release DNA
2. Purify DNA through precipitation with alcohol and salt 3. Use a spooling rod to separate the DNA from the solution |
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What is E. Coli DNA like?
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It is a circular molecule with 3,000,000 base pairs that attach to the cell wall at several points
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Describe the procedure to isolate Bacterial Chromosomal DNA:
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1. Mix with EDTA
2. Add SDS 3. Add RNase 4. Add protease |
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What does EDTA do?
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EDTA forms chelates (complexes) with several kinds of metal ions that protects DNA from DNase degradation. DNases need Cations (Mg++) in order to digest enzymes. EDTA blocks the DNA from degradation because Mg++ cannot by digested by the enzymes.
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What does RNase do?
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RNase digests any unwanted RNA
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What does SDS do?
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SDS denatures proteins and dissolves cell membranes
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What does protease do?
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Protease is an enzyme that digests proteins.
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What are plasmids?
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extrachromosomal, doublec-stranded circular DNA molecules containing 1,000 to 10,000 base pairs (small)
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How do plasmids replicate?
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They replicate independently of chromosomal DNA
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What are vectors?
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Plasmids used for propagating foreign genes in molecular cloning
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What is Form I DNA?
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Supercoiled plasmid DNA
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What does DNA Gyrases do?
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It is an enzyme that causes DNA to supercoil
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What enzyme causes DNA to uncoil?
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DNA relaxing enzyme
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