Short PCR product

Alex Brands abbrands at artsci.wustl.edu
Mon Jan 7 13:46:03 EST 2002


On Sat, 5 Jan 2002, aes wrote:

> Hi.
> I am going to run PCR with 35 bp long primers, with an amplicon in between
> of 15 bp, so that the total lenght of the PCR product is 85 bp (for
> quantitation by real time pcr and sybr green1). My question is, what is the
> shortest posible amplicon? And is 15 bp to short?
> Thank you and happy new year

Have you considered synthesizing the amplicon?  you can design a handful
of overlapping oligos like this:

---------  ---------  ----------  -----------
-------------  ------------  -------  -------

Kinase the oligos, mix them all together and treat with ligase.





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