Biology Reference
In-Depth Information
Figure 5.1
Outline of a simple cloning project involving insertion of foreign DNA into a plasmid
vector, and its subsequent insertion into a bacterial host,
E. coli
, to produce multiple copies (clones)
of the foreign DNA in recombinant plasmids. Both foreign and plasmid DNA is cut with a restric-
tion endonuclease to produce ends that will allow annealing of the plasmid and foreign DNA frag-
ments. The addition of DNA ligase combines the two DNA molecules and the plasmid is inserted
into
E. coli
where it will produce multiple copies (clones) of the new, recombinant DNA molecule.
Subsequently, the plasmids will be extracted from their host cells and used for other studies.
described provide the theoretical background to many of the current kit pro-
tocols, although companies may change details or may not explain the steps or
chemical components for proprietary reasons.
5.2.1 The Pros and Cons of Kits
Understanding the theoretical bases of the techniques described in this chapter
provides a background helpful in understanding the procedures one carries out
