One+of+the+first+attempts+around+8-25-10

One of the first times this method was attempted was during these experiments additional pcr cycles for troublesome primer pairs 8-27-10 pcr to amplify genes with difficult primers 8-27-10 (experiment seems to actually have been started around 8-25-10)

In this experiment 2.5 uL of 2.5 pmol/uL oligonucleotide primer of each primer was added to a final solution volume of 50 uL for 2.5*"uL"*2.5*"pmol"/"uL"/(50*"uL") = 0.125*"pmol"/"uL" final concentration of primer or 2.5*"uL"*2.5*"pmol"/"uL" = 6.25*"pmol" total. This reaction was performed with dynazyme with PCR protocol "Amp of Ab Genes 2" with 30 cycles. What does this translate to with the more conventional uM representation? 2.5*"pmol"/"uL"*1e6*"uL"/"L"*1*"umol"/(1e6*"pmol") = (2.5*"umol")/"L" 2.5*"uL"*2.5*"uM"/(50*"uL") = 0.125*"uM" final conc.

Later I added another 1 uL of 2.5 uM oligonucleotide primer each (just in the 1 uL volume), 0.1 uL dynazyme, and I performed 10 more cycles. The final concentration of each primer would have been

(2.5*"uL"*2.5*"uM"+1*"uL"*2.5*"uM")/(50*"uL") = 0.175*"uM" What's a typical final primer concentration? About 0.2-1 uM according to http://www.scientistsolutions.com/t8484-whats+a+good,+typical+final+primer+conc_.html