Unformatted text preview: BSC 2010L - BACTERIAL TRANSFORMATION TRANSFORMATION Plasmids Plasmids
• Bacteria have extra genetic material Bacteria plasmids. plasmids
– Plasmids are replicated during cell division. – The genes found on plasmids are The expressed, giving that bacterium additional characteristics. Plasmids typically have only 1, 2 or 3 genes. only Plasmids Plasmids Restriction enzymes cleave DNA Restriction Bacterial Transformation Today’s Lab
1. Digest plasmids 2. Bacterial transformation Bacterial Transformation lab exercises exercises
We do 2 basic exercises in this lab: We
1. A restriction enzyme digestion of a bacterial plasmid.
• Compare 3 digested (B,E &H) and 1 undigested plasmids with gel electrophoresis 2. Bacterial transformation: introduce plasmids with antibiotic 2. ntroduce resistance to E. coli. E.
• Plate bacterial cells onto agar with ampicillin to screen for Plate transformed cells Where do we digest the plasmid? 2. Bacterial Transformation 2.
• E. coli - not resistant to antibiotics. • Bacterial cells are competent, i.e., they are ready to Bacterial take up extraneous plasmids. take • Plasmids - with a gene giving bacteria resistance to ampicillin. ampicillin • Once plasmids are internalized, their genes are Once expressed by the bacterium - antibiotic resistance = transformation. transformation Write an essay about this lab (5 points) Write • The purpose of these two experiments • Methods including the strategy of a control Methods in both experiments in • What are your predictions and results • One page, single spaced, Ariel 12pt One Complete the Drosophila prelab handout before Drosophila the next lab. the ...
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