help with ligation

Jil Tardiff jtardiff at westnet.com
Sat Apr 13 20:13:19 EST 1996


Regarding your ligation problems.....how long and under what conditions are
you treating with CIAP? In my experience, if you let the reaction go on for
too long the ends can be damaged. I may be a bit paranoid ;-), but I 
routinely do my CIAP treatments in restriction buffer for only 5-10 min at
37 deg C and follow with 15 min at 65 deg C. I usually use CIAP from Boehringer-
Manheim.

A longer ligation time *may* help (maybe 4 hours) but I doubt that this
is the major issue.

good luck,

 Jil Tardiff
 Albert Einstein College of Medicine 



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