Which DNA restriction enzym must I juse

Chris LaRosa clarosa at biocomp.unl.edu
Fri Mar 17 14:53:28 EST 2000


Just order primers for the region to be sequenced.  PCR the fragment of
interest.  Clean it up (Prep a gene, Gibco Concert PCR kit) Sequence
from the pcr fragment.   We do this routinely for hundreds of different
pcr products. 

Paul Helvensteyn wrote:
> 
> I have Sequence project at school. The meaning is that I have to
> sequence a pice of Lambda DNA this pice must be as long as 0,5 KB. But
> the problem is that I first have to cut my DNA with a restriction enzym.
> The place where it must cut is at de site 7131 and 7591. between these
> no cut must be placed. One more thing is that the enzym must not cut
> more than maximum 15 to 20 times, because the pice is isolated from a
> agarose gel, to be cycle sequenced with a fluorcentprimer with the
> A-symetrical sanger PCR-methode.
> 
> I hope someone can give me the answer to my question
> 
> W.Helvensteyn
> The Netherlands




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