Question

2. How did specific aspects of the procedures performed in this laboratory support your goals and/or reinforce the introductory information given at the beginning of this chapter?

3. Growth on which plate(s) demonstrates that you were successful in your transformation of wild type E. coli to ampicillin-resistant E. coli?

4. It was to be expected that colonies on the LB/amp +pGLO plate should be white while those on the LB/amp/ara +pGLO plate should be green. Explain why this is so.

5 You may have noticed small white colonies surrounding the green colonies on the LB/amp/ara +pGLO plate. Explain why you think these colonies are present. Have these cells been transformed or not? Design an experiment that would allow you to demonstrate your hypothesis.

6. If you did the activity on transformation efficiency, explain why you think transformation efficiency varied between the groups in your class.

Answer 1

3.

Growth on LB containing Ampicillin demonstrate that tou are successful in your transformation.

4.

This is because, the Gfp gene in the pGLO plasmid is under arabinose promoter. So when the arabinose is present in the medium, this gene will be expressed and produce Gfp protein which makes colonies appear green under UV light.

5.

Because Gfp protein is being synthesised in these colonies in the presence of arabinose. Yes these colonies are transformed. To confirm wether these colonies are transformed, you can do plasmid isolation from those colonies and run on the agarose gel.

6.

because concentration of reagents used in transformation, amount of DNA, Phase of the bacteria, number of bacterial cells, these are some of the fectors that affects transformation frequency. Among different groups there might be slightly difference in these factors.