Dephosphorylation

GIETZ at bldghsc.lan1.umanitoba.ca GIETZ at bldghsc.lan1.umanitoba.ca
Thu Mar 4 13:36:00 EST 1993


From: angelo at phoenix.Princeton.EDU (Angelo Gunasekera)
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we have successfully used Shrimp Alkaline Dephosphatase to
dephosphorylate 5' phosphate.  The best thing about it is that it can be
irreversibly denatured by heating the reaction mix. to 70c for few
minutes.  You don't even have to do a phenol extraction to remove the
SAP as it will all be inactive.  We buy SAP from USB.
Good Luck
Angelo
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Hi Angelo!
    I was wondering how effective is SAP?  What percentage
of the ends get phosphatased?  What percentage of non-inserted 
plasmids do you get if you are cloning a sticky end to a pUC like
vector? (Blues vs Whites) I am wondering if I should switch from 
CIAP from BM.
________________________________________
Dan Gietz
Department of Human Genetics
University of Manitoba
770 Bannatyne Ave, Rm 250
Winnipeg, Man, Canada
R3E 0W3
GIETZ at BLDGHSC.LAN1.UMANITOBA.CA
_________________________________________





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