To do so, match the outcomes with the error(s) causing each. Under each “match\"

Question

To do so, match the outcomes with the error(s) causing each. Under each “match" provide an explanation Each match is worth 1 point (1/2 point for the correct letter, 1/2 point for a reasonable, concise explanation) (assume that only one error occurred at a time. Also note that the plasmid, recombinant or otherwise, must be circular in order to be replicated) (only use one outcome per error; each outcome is used once) pMB 1ori HindIII Spht Pst pUC19 2.686 kb Smal Kpni Sact EcoR amp Ndel Scal Aatl Sspl Outcomes a. Only well-separated blue colonies result, no white colonies are produced. b. Only well-separated white colonies result, no blue colonies produced c. No colonies grew on the plate d. Plate is covered in a bacterial lawn of white cells. (may be some blue cells but they are really rare) e. Plate is covered in a bacterial lawn of blue cells Errors A Forgot to add X-Gal to the agar growth medium in the plate Used a strain of bacteria that expressed its own, genomic copy of beta-galactosidase (usually a strain with a nonfunctional copy is used) Added the ampicillin to the growth medium too early (before the medium was allowed to cool), thereby destroying the effectiveness of the antibiotic (ampicillin doesn't work) Mistakenly heated up the tube of ligase enzyme (cost of $137) in the water bath, inactivating it before using it in the ligation reaction Used a strain of bacteria in the transformation step that already carried the pUC19 vector

Explanation / Answer

Forgot to add X-gal to the growth medium in the plate -Outcome B. Lac-Z degrades X-gal into a blue color compound. So when the bacteria transformed with the pUC vector will be able to cleave the X-gal and gives blue color colonies. Since X-gal is not added to the plate so we will get all white color colony.

Error B- Outcome A., In this case, bacterial cells will make beta-galactosidase and all the cells will be able to cleave X-gal and we will get all the blue colonies on the plate

Error C- Outcome D. addition of ampicillin in the growth media when the media is still hot leads to degradation of ampicillin. Since there is no antibiotic so all the bacterial cells present in the competent cells vial will grow on the plate.

Error D - outcome C. heating the ligase enzyme will compromise its activity as a result of it enzyme will not be able to ligate the DNA fragments. SO we will not get any colony on the plate.

Error E- Outcome E- Since the bacterial cells already have the pUC19 vector so all the cells will produce Beta-galactosidase which will give us blue colonies on the plate

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