complete removing of primer

Rafael Maldonado rafael at howard.genetics.utah.edu
Fri Feb 16 17:26:52 EST 1996


On Fri, 16 Feb 1996, Kyung-Kwon Lee wrote:

> I am trying to remove primers(30mer) completely(more than 1000 fold)
> before 2nd PCR with other pirmers.
> A tiny amount of pirmers remained causes nearly no amplication products of
> 2nd PCR. Change of primer sequence is desirable but limited sequence
> informations don't allow any alternatives.
> Unfortunately,the only way is to remove first primers compeletely.
> Please any adivces and experiences on methods removing primers completely.

I have used succefully those methods:

- Gel purification of the amplified DNA fragment. Cut the band and 
extract the DNA with any method described elsewhere.

- Use Centricon 100 (if you fragment is bigger than 10 pb). Wash several 
times (5 X 2 ml) and the oligos won't mess in the next PR.


Good luck


Rafa

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