PCR primers for common plasmids

John Watson watson_j at bms.com
Tue Oct 22 17:25:53 EST 1996


Ethan Strauss wrote:
>
> I have been getting clones from the Arabidopsis EST bank. I would
> like to be able to amplify the inserts using primers directed against
> the vector. Most of these clones are in lambda ZipLox which has T7 and
> SP6 RNA polymerase transcription sites and M13 forward and reverse
> sites.
>
> Can anyone give me advice on primer design for PCR using these
> sites? Will the standard sequencing primers sequences work?
> If I do go with sequencing primers is it always cheaper to custom order
> them to buy them as sequencing primers? Why?

I always use just the standard sequencing primers, although you may find 
that the Tm for some of them is a little on the low side.  Still, given the 
low complexity of the average miniprep, this is not usually a problem.  I 
have also synthesized slightly longer versions of these primers, but I do 
not think that it is really necessary.  

I have no idea which is cheaper -- I always use in-house oligonucletide 
synthesis.
----------------
John Watson
Bristol-Myers Squibb Co.
watson_j at bms.com
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