One of the first attempts around 8-25-10

2015-01-13

azim58 - 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

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