Single primer PCR amplification??
landel at helios
Fri Jan 27 11:09:31 EST 1995
In article <D30yJs.E1F at info.swan.ac.uk>, <bajenkins at swansea.ac.uk> wrote:
>In article <1995Jan20.210640.14826 at emba.uvm.edu>, edahl at moose.uvm.edu (Eric N. Dahl) says:
>>This may be a stupid question... is it possible to amplify DNA using just
>>one primer? How difficult is it to get it to work?
>>One primer PCR is certainly possible and is quoted in many
>places as a good way of producing single stranded template
>You must however start with a PCR product, and use quite a bit of it
>(1-10%), this ensures that the Taq doesn't extend your primer
>indefinately, e.g if you used genomic DNA.
>The amplification will not be exponential, but linear for obvious
>reasons, hence the requirement for sufficient template.
Check out this paper:
1. Weis JH.
'Race no more': an alternative approach to cloning the 5' end of
Nucleic Acids Research, 1994 Aug 25, 22(16):3427-8.
The author reports that he got 5' RACE to work using only the internal
primer. The hypothesis is that at some low frequency, the primer somehow
gets incorporated into both ends, but when that happens, amplification
goes as normal.
I thought it sounded far-fetched, UNTIL
We were doing some 5' RACE, and sure enough, we got some products with
the same primer on both ends!
More information about the Methods