You might just try calculating the Henderson hasselbach calculation using activity coefficients, which takes into account the ionic strength of the solution.
Doing this with pH calculations makes a decent amount of difference.
I was writing some python code to do this calculation, lemme go finish it up.
On Feb 6, 2013 6:28 PM, "Josh Perfetto" <josh@snowrise.com> wrote:
-- Hi,I'm wondering what the expected accuracy of UV spectrophotometers are when quantifying dsDNA. I had DNA in TE which I believed was 100 mg/ml based on resuspending a known quantity of DNA as measured by a synthesis company. I dilluted it 50x to a total volume of 100ul and measured in two different spectrophotometers.Spec 1 (Thermo Electron Helios Gamma):A260 = 0.031 AA280 = 0.019 ACalculated concentration: 77.5 ug/ml--Spec 2 (Eppendorf BioPhotometer):A260 = 0.050 AA280 = 0.022 ACalculated concentration: 125 ug/mlThe consistency was a little disappointing. Is this normal or do you think there is a problem with my specs? I suppose I could get more accuracy by diluting less but I hate using up so much plasmid just for quantification.-Josh
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