Direct Selection and Clone Identification Cloning Strategies
There are two strategies for obtaining the clone
Ilustration of the problem of selection
Although there are many different procedures
by which the desired clone can be obtained, all are
variations of two basic themes.
- Direct Selection for the desired gene, which
means that the cloning experiment is designed in such
a way that the only clones that are obtained are clones
of the required genes. Almost invariably, selection
occurs at the plating out stage.
- Identification of the clone from a gene library,
which entails an initial 'shotgun' cloning experiment,
to produce a clone library representing all or most
of the genes present in the cell, followed by analysis
of the individual clones to identify the correct one.
In general terms, Direct Selection is the preferred
method, as it is quick and usually unambiguous. However,
it is not applicable to all genes.
Techniques for clone
identification are therefore very important, especially
as complete genomic libraries of many organisms are
This guide will explain on how to perform the cloning
using the Direct Selection method.
Cloning by using the Direct Selection Method...