Worksheet 13 (answers might be incorrect) PDF

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Worksheet 13 (personal answers)...

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Name: Meet Patel

Section:

014

Worksheet 13: Transformation Below are results from a transformation based on the procedure you had conducted. Using the results below answer question 1.

1.

Plate

No Plasmid

+ Plasmid

LB plate

Hundreds of colonies

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LB/ampicillin plate

no colonies

20 colonies

Was the transformation successful? Explain. (1.0 pt)

Yes, the transformation was successful. We saw the expected results for E. coli (No plasmid, LB plate) to see hundreds of colonies. For E. coli (No plasmid, LB/ampicillin plate) we saw no growth on the plate which was due to no presence of plasmid. For E. coli (+ plasmid, LB/ampicillin plate) we saw growth on the plate which was due to presence of plasmid. Thus, the transformation of E.coli was successful since with the presence of plasmid we observed the growth on LB/ ampicillin plate. 2.

Clumsy McForgetful used the same plasmid and LB/amp plates as you used in lab, but his transformation was unsuccessful. What could be one reason Clumsy’s transformation did not work? (1.0 pt)

1. Not performing heat shock or incorrectly performing heat shock. 2. The temperature and length of the incubation period. 3. Improper use of cell-spreading tools.

3.

What was the purpose of plating the “no plasmid” condition on the LB/ampicillin plate? Is it considered a positive control or a negative control? (1.0 pt)

E. coli with “No plasmid” condition on the LB/ ampicillin plate: It acts as a control to confirm that bacteria cannot grow in the presence of ampicillin. It is considered a negative control. Ampicillin is an antibiotic and works by preventing E. coli from constructing cell walls, thereby killing the bacteria. If the bacteria are viable on the LB/amp plate, then they are resistant to ampicillin. If no bacterial colonies survive, then they were not ampicillin resistant (they were ampicillin sensitive)....