inosine in PCR primers

Dr Richard Sallie sallier at
Thu Jun 27 08:24:14 EST 1996

There has been quite a bit published on this, mostly in Nucleic Acids
Research.  One of the important issues is what the 4th position will be
used for: If it contains a RE site that will be important to yo later,
then use of inosine may be a problem, depending on the enzyme to be used.
Otherwise, it is generally OK to use inosine in this position.
Richard Sallie
In article <4qrlhr$3ok at>, bapn4 at
(Martin Goodson) wrote:

> Dear all,
> In order to design a set of PCR primers I need to know how inosine affects
> the binding and Tm of oligos. For instance, is it OK to have an
> inosine 4 bases away from the 3' end of the primer or should this
> position be kept four-way degenerate? Has there been anything published
> on this?
> thanks in advance
> martin g

Richard Sallie MD FRACP
Consultant Physician and Hepatologist,
Perth, Western Australia

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