Does sodium citrate inhibit RT-PCR?

Alan Smith alansmith at students.wisc.edu
Wed Oct 10 17:12:10 EST 2001


Hello,

Will a solution of 1 mM sodium citrate pH 6.4 inhibit either the cDNA
synthesis step or the PCR step of an RT-PCR reaction?  I understand that
sodium citrate can chelate metal ions and this may be the problem. Recently
our RT-PCR reactions have stopped working.  Could it be caused by performing
the reactions in a solution of mainly 1mM sodium citrate? I read in a qiagen
tech help manual that 1mM sodium citrate does not inhibit PCR reactions, I
could not find any reference to RT-PCR.  Any help on this topic would be
appreciated.  If it does inhibit RT-PCR how much Mg should be added to a
20ul reaction to prevent inhibition?

Thank You

Alan Smith

Graduate Student
Plant Breeding and Plant Genetics Program
University of Wisconsin-Madison

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