RT reagents affect PCR

Steve Hopkins shopkins at fs1.ho.man.ac.uk
Thu Dec 8 22:59:22 EST 1994


While developing a quantitative PCR method we have become aware that transfer
of small amounts of the cocktail of reagents in the reverse transcriptase
reaction (many of which which are present at much higher concentration than
those in the PCR-mix) influences the subsequent PCR. This means that the PCR
signal may be a function of the amount of RT-mix transferred, as much as the
amount of cDNA. It could also be particularly important where cDNA standards, 
not in RT-mix cocktail, are used. Has anyone experience of what RT-mix factors 
might be important? Can transfer of random primers affect PCR? We are checking 
some of these things out, but any help would be appreciated.




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