In producing a recombinant plasmid to be used to clone a given donor insert, it is possible to cut both the donor and plasmid with the same restriction enzyme, resulting in complementary sticky ends. Assuming plenty of plasmid DNA is available, why is further selection necessary before the introduction of the plasmid into a cellular system?
A)Contamination will have introduced other donor inserts.
B)Some donor inserts will be single stranded and deteriorate.
C)Some donor inserts will be sensitive to particular antibiotics.
D)Some donor pieces will remain uninserted.
E)Some donor strands will be inserted with an incorrect orientation.

Question

Quizplus · Accepted Answer

Even if the donor insert and plasmid are cut with the same restriction enzyme and have complementary sticky ends, there is still a possibility that the donor insert can be inserted into the plasmid in the wrong orientation. This would result in the incorrect expression of the gene of interest or complete inactivation of the gene. Therefore, further selection is necessary to ensure that only recombinant plasmids with the donor insert in the correct orientation are introduced into a cellular system.

In Producing a Recombinant Plasmid to Be Used to Clone