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Molecular Biology, 4/e
Robert F. Weaver

Molecular Cloning Methods

Steps in Cloning a Gene

View the animation below, then complete the quiz to test your knowledge of the concept.



1

Arrange the following events in the proper sequence for gene cloning.
1 = Incorporate gene into bacterial plasmid
2 = Isolate DNA from organism containing desired gene
3 = Incorporate cloned gene into bacterial cells
4 = Fragment DNA with restriction enzyme
A)1, 2, 3, 4
B)2, 1, 4, 3
C)2, 3, 4, 1
D)2, 4, 1, 3
E)2, 4, 3, 1
2

The function of the enzyme ligase is to
A)add new DNA to the plasmid.
B)remove DNA from the plasmid.
C)form covalent bonds between cloned gene and plasmid.
D)repair damaged DNA.
E)purify the plasmid DNA.
3

"Sticky ends" are
A)single-stranded DNA sequences that are generated by staggered cuts.
B)double-stranded DNA sequences that are generated by staggered cuts.
C)single-stranded DNA sequences that are generated by blunt cuts.
D)double-stranded DNA sequences that are generated by blunt cuts.
E)different from cohesive ends.
4

The plasmid used for cloning has a single restriction site.
A)True
B)False
5

Restriction enzymes cut DNA at random sites.
A)True
B)False