Several lab groups independently carried out similar ligation/transformation steps that you
pe...
Several lab groups independently carried out similar ligation/transformation steps that you
performed in the lab to introduce GFP into the MCS of the phosphatase treated digested pUC
vector. Both the vector and GFP sequence had been digested with the same single enzyme. The
results obtained by each of the groups were as follows:
# of colonies on LB-Amp plates
Transformation into XL-1 Group 1 Group 2 Group 3 Group 4
a) Cut pUC + GFP insert 0 529 2 1125
b) Cut pUC 0 2 5 930
c) GFP alone 0 0 0 890
d) 10 uL uncut pUC (0.5 ng/uL) 0 975 975 975
e) No DNA 0 0 0 925
6. Which group obtained the expected/desired pattern of results?
7. Which group would be expected to give the greatest percentage of blue colonies on X-Gal
plates with the transformation mixture "a"?
8. Which transformation mixture would be expected to have the highest percentage of
intramolecular ligation events?
9. What was the transformation efficiency of the competent XL-1 cells? Indicate the number of
transformants expected per microgram of undigested DNA.
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