Transformation-1101 - Biotechnology/Molecular Biology...

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Unformatted text preview: Biotechnology/Molecular Biology Bacterial Transformation Biotechnology Branch of biology dealing with the study of DNA and genetics on the molecular level Biotechnologists manipulate DNA and genetic information to produce particular products or to study genetic disorders or diseases Medicine, agriculture, genetics, and the food industry Biotechnology Recombinant Technology Transformation: the placing of new genetic material into a cell changing the genotype and possibly the phenotype Vectors: entities that carry foreign DNA into the cell of an organism where DNA is expressed We will use a plasmid that confers antibiotic resistance to the transformed organism - E. coli cells Sterile Technique Don't open sterile packages until ready for use Avoid touching tip of pipet/transfer loop to anything Don't reuse tips prior to resterilization Keep sterile liquids covered Flame spreader and loop before/after each use Open petri dish only as needed, with it inverted when possible Deoxyribonucleic Acid http://www.accessexcellence.org/AB/GG/dna_molecule.html E. coli A gram-negative rod shaped bacteria found in the human intestine Has a single circular chromosome Can be easily cultivated and studied Grows rapidly and is highly used in lab settings Electroporation A small amount of electricity is applied to the cells in the presence of the plasmid This will create transient holes in the membrane which enable the cell to take up new DNA Cells that contain the plasmid (transformed) will be able to grow in the presence of ampicillin Micropipettes These tools can be used to accurately measure small amounts of liquid samples They come in a range of sizes and use tips, which provide sterility and prevent contamination Using Plasmids in Science Restriction Endonucleases Recognize a specific sequence and cleave the phosphodiester backbone of both strands of DNA at or near the recognition sequence Sequences are often palindromic ...
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This note was uploaded on 05/02/2008 for the course BIOL 1101 taught by Professor Holmes during the Spring '08 term at East Carolina University .

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