PCR & spermidine

hodges bradley l segdoh at uxa.cso.uiuc.edu
Fri Feb 10 15:34:37 EST 1995


You didn't mention if there is an intron(s) between your primer pairs on 
the genomic DNA.  TAq polymerase can PCR 2kb at the most (in my 
experience).  So if this true that may be your problem.  You can do long 
PCR by using TaKaRa LA PCR kit (up tp 20-30kb).


On Thu, 9 Feb 1995, Claus B. Jorgensen wrote:

> Hi Bionetters,
> 
> I have a couple of primer pairs that works nicely on the plamid template, but 
> they fail when I'm turning to genomic DNA. 
> I have tried different [Mg] and annealing temps, but neither seems to change 
> the outcome.
> 
> Any suggestions to modifications would be appreciated. 
> 
> BTW. I seem to recall a question some time ago about spermidine in PCR 
> reactions and the wonders it could make. If anyone has got any experience I 
> would be pleased to hear.
> 
> Cheers,
> 
> Claus B. Jorgensen
> Animal Genetics
> RVAU, Copenhagen
> Denmark
> E-mail: cljoh3 at wptemp.kvl.dk
> 
> 
> 



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