Another attempt around 4-4-13

2014-08-29

azim58 - Another attempt around 4-4-13

In this reaction PrimeSTAR Max was used up to a 50 uL total volume. 1 uL
of 2 uM of each primer was used for a final conc. of
1*"uL"*2*"uM"/(50*"uL") = 0.04*"uM". 30 cycles were performed.
Later I added 1 uL of 10 uM primer and added 5 uL primestar max premix as
well as 5 uL H20 and performed 5 more cycles
Another 1 uL of 10 uM primer was added and 5 uL primestar max premix as
well as 5 uL H20 was added and 5 more cycles were performed.
Final concentration of primer would have been
(1*"uL"*2*"uM"+1*"uL"*10*"uM"+1*"uL"*10*"uM")/(50*"uL") = 0.44 uM

What if I had not done 2 rounds of 1 uL 10 uM addition? Then what would
the primer concentration have been?
(1*"uL"*2*"uM"+1*"uL"*10*"uM")/(50.0*"uL")=0.24 uM which would have been
quite reasonable.
^What if I just added 0.5 uL of 10 uM primer?
(1*"uL"*2*"uM"+0.5*"uL"*10*"uM")/(50.0*"uL") = 0.14*"uM" which is still
reasonable
^What if I want to end with a final of 0.3 uM?

(1*"uL"*2*"uM"+y*10*"uM")/(60.0*"uL") = 0.3*"uM" -> y=1.6 uL

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