PCR Self Adjusting Magnesium concentration

tiredkhan at yahoo.com tiredkhan at yahoo.com
Thu Jun 29 17:19:05 EST 2006


The Hotmaster PCR mix from Eppendorf contains a weak chelator of
magnesium that is claimed to result in just the right amount of free
Magnesium in the PCR so that there is no need for magnesium titration.

I was just wondering if anyone who has used this mix could give
feedback about this - do you feel from your experience that this might
be true?

If I have one reaction that only works well with very low free
magnesium concentration such as 0.3 mM, and another that works better
with 2 mM free magnesium, will both work just as well with a self
adjusting mg buffer? I guess it sounds unlikely to me so I was inclined
not to spend and do the experiments, but if there are people out there
who are very enthusiastic about this type of approach then maybe I
will.

Otherwise, does anybody have ideas as to what weak chelators to add to
our usual PCRs, or references to work on metal ion buffers - I haven't
been able to find anything useful in my searches.

Many thanks, TK



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