IUBio

DNA ligation help

Nick Jacobsen jacobsen at cf.ac.uk
Mon Jun 3 04:20:12 EST 1996


Hi,
I find that the majority of problems with DNA ligation inefficiency 
are due to ATP degradation in the reaction buffer. If in doubt order 
new buffer and freeze in small aliquots. You can also order Pharmacia 
ligase - you have to order separate 10mM ATP with it to add to the 
reaction as required. You have to be fairly accurate with the amount 
of ATP added as too much is as bad as too little.

I hope this is of help to you

cheers

Nick Jacobsen.




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