Digestion problems with EcoRI

M.S.Rodriguez Pena m.s.rodriguezpena at amc.uva.nl
Wed Nov 21 06:31:54 EST 2001


Hi Barbara,
look in the new NEB catalog (2000-2001) pag 214. They mention site
preferences for some restrictions enzymes (Eco RI among them). You might
have the same problems I had with NaeI. After one week of dayly digestions
I could not figure out why NaeI was not able to cut my plasmid.


Marisol

Barbara Simionati wrote:

> Hi,
>
> I am trying to double digest salmonella genome with EcoRI and MseI but
> while MseI works (complete digestion) EcoRI doesn't.
> I can't understand why.
> I have tryed enzimes from two different tubes
> I have tried up to 80 U
> In both cases I could see the undigested genomic DNA
> I have digested a pGem and it worked.
>
> DNA was prepared with a Qiagen Kit
>
> Thanks for any suggestions
>
> Barbara Simionati
> barbara at cribi.unipd.it
>
> University of padova
> Italy
>
> ---

--
Dr MS Rodriguez Pena, MD, phD
Laboratory for Experimental Internal Medicine
Rm G2-136
AMC. Meibergdreef 9
PO Box 22700
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Tel 31-20-5662368
Fax 31-20-6977192
e-mail: m.s.rodriguezpena at amc.uva.nl





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