Help with a knotty ligation?

Cody Buchmann buchmann.3 at osu.edu
Tue Dec 18 18:14:06 EST 2001


1) Yes, I am, and I usually run a separate ligation/transformation control
with untreated vector to see how efficiently the CIAP works.

2) "Prophylactic cutting" is something we've done in the past to reduce
background; it's a very good idea.  Unfortunately it doesn't do much in this
case; the background is very low, but there is no foreground whatsoever. 
We've shown that the ligation/transformation isn't just inefficient, it's
entirely zero.

3) I'm definitely going to suggest this to my PI.  (Another suggestion along
these lines involved ultra-slow ligation...ligating at 4 degrees for 48
hours).  Does this refer to the SOC stage after heat transformation (we
don't normally do electroporation) or incubating the plates after
transformation?

-- Cody Buchmann
   Children's Hospital, Columbus, OH


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