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.



More information about the Methods mailing list