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In the first activity, students use a rapid method to render the bacterium Escherichia coli "competent" to uptake plasmid DNA.
Member rating: No Rating | Words: | Submitted: Wed Aug 13 2003
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In the first activity, students use a rapid method to render the bacterium Escherichia coli "competent" to uptake plasmid DNA. Escherichia coli cells are then transformed with the pAMP plasmid, which carries the gene coding for resistance to ampicillin. The recipient E. coli strain, MM294, lacks this gene; thus only transformed cells acquire the ability to grow on ampicillin. Ampicillin then is the selective agent in this experiment. E. coli cells are scraped from two large colony starter plates and suspended in two tubes containing a solution of calcium chloride. pAMP plasmid is added to ONE cell suspension, and both tubes are incubated at 0 deg C for 15 minutes. Following a brief heat shock at 42deg C, cooling, and the addition of LB broth, samples of the cell suspension are plated on two types of media: plain LB agar and LB agar with ampicillin. The plates are incubated for...
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