Quick question: splitting TOPO cloning kit - (Jun/28/2006 )
I was wondering if anyone has had success with using a 20 rxn Invitrogen cloning kit for 40 rxns?
trying to squeeze more out of this kit....
Absolutely. It's routine.
Yeah, I have a tendency of ignoring Invitrogen's "recommendations."
Yes, I have done a lot for saving money for my boss although he did not increase my salary...
haha- I too noticed that saving money for the lab does not translate into higher wages....
Thanks again to everyone who responded to my question.....
In order to save more money, why don't you try to do MaxiPrep of your vector? Then you will be able to do hundred of reactions (the efficiency will probably not be so high as the original ones but it will be good enough).
Do many people do this? I believe that it is a common practice. Can anyone share their experience to this respect?
The Topo vectors have been treated with vaccinia topisomerase, which binds to CCCTT sequences and mediates the insertion with high efficiency. A maxiprep will not reproduce the efficiency of topo cloning reactions. There are commercial sources of vaccinia topoisomerase, however.
Phage is correct.
TOPO system they provide is linearized and can't be propagated.
That doesn't mean you have to use TOPO, however...most of the time I digest PCR product and clone right away into the desired vector.