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>
writes
>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.

Duncan 
--------------------------------------------------------------------------------
The problem with being on the cutting edge is that you occasionally get 
sliced from time to time....
http://www.thenet.co.uk/~dnamp



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