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Lab report 7 - Plasmid Lab Lab 7 Christopher Chu Wednesday...

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Plasmid Lab Lab 7 Christopher Chu Wednesday 6:00 – 10:00 pm Objectives A) To transform bacterial cells so they can express a new gene by inserting foreign DNA into the cells via a plasmid transformation. B) To detect the expression of the new gene by screening for the bacteria C) To learn techniques for handling and culturing bacteria D) To observe the expression patterns of genes involved in epidermal cell-fate determination in Arabidopsis. E) To observe and recognize several different phenotypes of Arabidopsis mutants. Materials Arabidopsis, different types of plasmid, E. coli, ice bath, warm water bath, and spectrometer, X-Gluc, ethanol, paper towels, permanent marker, microfuge tubes, micropipetter, centrifuge, agar plates, bacterial spreaders, rubber bands, containers to dispose of contaminated material. Procedure We began the plasmid lab by squirting ethanol on our work place to make it sterile. Keeping the E. coli on ice, we mixed the E. coli with the different plasmids, and kept one
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tube of E. coli unchanged. This was to be our control. Next, we placed the mixtures of plasmids and bacteria into a warm water bath which provided the heat shock that would allow for the transformation. We plated these bacteria into plates containing either agar or agar with ampicillin and used a spreader to spread the bacteria across the entire plate. The next week, we observed the effects of the plasmids and the results of the transformations.
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