Streptavidin and Biotin interactions
tguettou at newssun.med.miami.edu
Mon Feb 10 16:37:40 EST 1997
On 9 Feb 1997, Diane Webster wrote:
> I am currently using biotin labelled oligo(dT) to capture my
> polyA tail potyvirus in streptavidin coated tubes for RT-PCR
> (Boehringer Mannheim's mRNA capture kit).
> I would like to know more about how streptavidin and biotin interact.
> Specifically do they dissociate when heated during PCR (94 C)?
> And does the oligo(dT) dissociate from the biotin label during PCR?
> General related details on streptavidin and biotin would be useful
> Thanks in advance
> Diane at phyton.otago.ac.nz
> Botany Department
> University of Otago
> PO Box 56
> New Zealand
The interaction between Biotin (Vitamin) and Avidin (protein) is one of
the strongest known in biological systems. It is stronger than any
antibody/antigen interaction. You are using streptavidin which lacks a
glycosylation site thereby decreasing nonspecific binding. Since avidin
is a protein, it will be denatured during PCR and therefore release
biotin. I don't know if it will bind Biotin again, when you lower the
temperature during the cycle, but I doubt it. Biotin is fairly stable and
since it is covalently attached to your oligo it should not come off. You
can also check its temp stability in the Merck index or call the company
you bought it from.
BTW there is a lot of literature about this out there. A lot of companies
have handbooks describing the interaction and properties (e.g. Pierce).
Hope that helps
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