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Cloning Materials and Methods
The basic methodology involved in gene cloning is summarised as follow
a fragment of DNA you want to clone is introduced into a structure
called a "cloning vector" to produce
a recombinant DNA molecule (figure 1). The vector is a fragment of DNA, containing genes able to ensure its own replication within a host cell.
The recombinant DNA molecule is introduced into a host
cell by transformation (figure 2).
Within the cell, the vector's genes will direct the multiplication of
the recombinant DNA. This cell will start making copies of the vector + insert.
When the cell divides, copies of the recombinant molecule are passed on
to the daughter cells of the bacterium.
A large number of cell divisions gives rise to a clone. In
a clone, all the cells are identical, and come from the same initial cell.
They all contain copies of the recombinant DNA molecule.
Cloning DNA is, in theory, a very straightforward procedure. Its success is
due to the fact that cloning can isolate genes, or fragments of genes from
the rest of the DNA. That is what happens when one is constructing a genetic bank.
The genome is cut into fragments and each fragment is introduced into a
vector (usually a virus). It is then possible to replicate the virus in great quantities and select a clone of the desired gene. The methods used differ, depending on the laboratory.
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