Pvu II partial digest conditions needed
Dr. Duncan Clark
duncan at genesys.demon.co.uk
Mon Mar 24 10:57:11 EST 1997
In article <obrien-2303971742090001 at brass6.med.upenn.edu>, ":-Peter"
<obrien at pharm.med.upenn.edu> writes
>Pvu II has STAR activity (per the NEB catalog), so I'm afraid to encourage
>low Pvu II activity with high DNA concentrations, using the wrong buffer
>to reduce activity etc. All I can think to do is to do the digest at room
>temp or colder. to get enough DNA to play with.
Star activity is the other way high enzyme, low DNA ie a very high
overdigest. Don't forget that Nae is very site selective on colE1 based
plasmids ie some sites cut 50 x slower than others. There is a 'Turbo'
version of Nae that gets around this I think from Promega.
The problem with being on the cutting edge is that you occasionally get
sliced from time to time....
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