How to pull-out a specific PCR product?

Yoram Gerchman yoramg2 at LEONARDO.LS.HUJI.AC.IL
Thu Feb 12 12:27:11 EST 1998


On Tue, 10 Feb 1998, Jill Kellogg wrote:
>
> Its not obvious to me what makes the primer that amplified your 
non-specific $
> products different from the primer that amplified your desired product.
Tag$
> the primer with something won't help you separate the non-specific from
the
> desired product if they are both amplified with the same primer   Nested
> primers, if you have enough data, are the way to go.

The thought was that a random primer might stick on allot of places (both
strands), so one will get both semi-specific products (one side specific)
and totally-unspecific products from the random primers alone. The tag will
enable enrichment of the semi-specific product. A nested PCR would have
been nice but we don't have enough data.

.........................................................................
Yoram Gerchman
Division of Microbial & Molecular Ecology
The Institute of Life Sciences
Hebrew University of Jerusalem
Givat Ram, Jerusalem 91904, Israel
Tel: 972-2-6585175




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