quick ligation?
TJ Murphy
medtjm at bimcore.emory.edu
Fri Jun 21 17:46:54 EST 1996
Alan Hair wrote:
>
> I have noticed various kits offering very fast ligation times. What is the
> magic ingredient and is it any good for ligation of double stranded
> linkers to DNA?
> Cheers
> Alan
Got me. If I ligate longer than 10 minutes with your standard
reagents it's usually because
A) I want to go home
B) I have to go home
C) I'm having difficulty scrounging up competent cells
Convince yourself. Take a PhiX/HaeIII ladder into two tubes. Add ligase
overnight for one, and for a few minutes for the other. Run the gel.
You'll see no difference. That is, of course, the painful blunt ended
ligation ;)
You can probably devise a similar experiment to test your linkers. Just
use the latter in molar excess, and if you run a high percentage gel far
enough, you'll see a slight shift in size of any small molecular weight
fragments in your ladder. If your ladder ligates up, you got a linker
problem.
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