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During our discussion of replication origins we took a step-by-step look at the method of cloning DNA fragments into plasmids. Based on your knowledge of this discussion which of the experimental manipulations would lead to the failure to insert a fragment of genomic DNA into the plasmid vector?

A. Digest plasmid DNA with the BamH1 restriction enzyme and genomic DNA with the EcoR1 restriction enzyme. 

B. Add bacteria that have been transformed with a plasmid having the kanamycin (antibiotic) resistance gene to an agar plate that having chloramphenicol (antibiotic).

C. DNA polymerase is not added to the ligation mixture.

D. Add bacteria that have been transformed with plasmid DNA to a plate that lacks an antibiotic.

 

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