AP Biology Bacterial Transformation What are each of

AP Biology Bacterial Transformation

What are each of the plates for? LB, LB/AMP • LB: This is the standard agar that all bacteria grow on • LB/AMP: This plate contains ampicillin, only bacteria that contain the gene for the enzyme betalactamase, which breaks down ampicillin, will grow on this plate , our plasmid contains the Betalactamase gene

What is the purpose of the –plasmid plates? • These are the controls. • The LB -plasmid plate will test to see if the bacteria can still grow after we’ve run tests on them • The LB/AMP -plasmid plate should show no growth because the bacteria have not been exposed to the plasmid with ampicillin resistance.

Plasmids • The plasmid has the antibiotic resistance gene and the p. VIB gene. Bacteria that accept this gene will grow on a plate with antibiotic in it. Bacteria without the plasmid will not grow on a plate that contains ampicillin.

What is a satellite colony? • Bacteria that are not transformed (have not taken in the plasmid) but are growing close to the colony that did because the transformed colony has destroyed the ampicillin around it. Satellite colonies Transformed Colony

What is the purpose of the Ca. Cl 2 and the glass beads? Ca. Cl 2 Glass Beads • Changes the polarity of the cell membrane of the E. coli so they more readily accept the plasmid. • Helps to spread out the bacteria on the plate

Sterile Technique • No food, drinks or gum in the lab at all! • All tables are washed with soap and dried at the end of the lab period. • All of us will wash our hands before we leave the lab. • Nothing can be set down on the lab table before or after it has touched bacteria. • All lab “trash” will go into the waste disposal bag provided. • This is not paper towel trash. Paper towels still go into the regular trash cans.

Ecoli w/ the PVIB plate