a) Describe how you could determine quantitate (determine how many) E. coli cells have been transformed?

b) Briefly outline your experiment. You will need a control, or standard, to analyze your experimental results, make sure to indicate this in your answer – explain what the control/standard is and how you use it.

Solutions

Expert Solution

Transfored Ecoli must be resisitant for the particular antibiotic. The steps are as follows

1. Say we need to transform E.coliBL21 with the plasmid DNA. This plasmid DNA contain antibioticresistant gene.

2. Tranform the bacteria as per protocol in the presence of antibiotic. So that only transform colonies will grow. Say culture volume is 5 ml.

3. take a 0.1 ml of the culture and streak it on the agar plate containg antibiotic. Incubate the plate overnight.

4. Count the colony number. Each colony is generated from sinle transfored cell. Say for e.g. 3 colony has been developed.

5. Therefore 0.1 ml of the culture contains 3 transformed cell.

6. Similarly it can be calculated for orignal 5 ml of culture.

(5*3)/0.1 = 150.

Therefore transformed cell number in the orignal 5 ml of culture after transformation is 150.

For this procedure there will be no need of use of any standard and control in the experimnet.

gladiator answered 1 year ago

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