NdeI: cutting problems?? anyone else?? seems silly!

Dr. Duncan Clark duncan at genesys.demon.co.uk
Sat Mar 9 08:18:10 EST 1996

In article <Pine.AUX.3.91.960308215654.7830A-
100000 at mail.med.cornell.edu>, Heidi Moss <hmmoss at MAIL.MED.CORNELL.EDU>
>greetings 'netters!!
>Has anyone else experienced problems with NdeI (ie. incomplete/non-cutting)?
>We have 2 lots from 2 different companies (BM and Gibco) and neither of 
>them are working. Trobleshooting (ie. altering buffer conditions--there 
>doesn't seem to be a consensus in the literature) has turned up nothing. 
>It is a mystery to us and the companies that supplied them. Even on 
>control DNA!!
>What is going on? Is there an NdeI god out there who has cast a spell? 
>All other enzymes are working perfectly.
>If you can shed any light on this problem, please let me know.
>thanks in advance!

Nde I is very sensitive to impurities in DNA preps ie minipreps cut very
poorly. It also has an extremely short half life of 15 mins. We do a lot
of cloning using Nde and use the NEB enzyme 'cos it is cloned and way
cheaper than alternative sources.

The problem with being on the cutting edge is that you occasionally get 
sliced from time to time....

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