Short PCR product

Duncan Clark junk at
Tue Jan 8 04:29:55 EST 2002

In message 
<Pine.GSO.4.31.0201071243480.19893-100000 at>, Alex 
Brands <abbrands at> writes on the Mon, 7 Jan 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.

But he's not trying to do gene synthesis.

He is running quantitative real-time PCR of an 85bp amplicon.

The problem with being on the cutting edge is that you occasionally get
sliced from time to time....

Duncan Clark
GeneSys Ltd.

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