One unit of DNA polymerase activity is conventionally defined as the amount of enzyme that will incorporate 10 nmol of nucleotides during a 30-min incubation. I found out about the fluorescence-based method. How do I calculate how many dNTPs have been incorporated?
I did find this paper but couldn't find the full text. Article Fluorescence-Based DNA Polymerase Assay
If anyone has a full text kindly do share.
Here it is - although it is quite old, you might want to check Google Scholar for newer publication on the topic
You could just buy a polymerase with known units and run it head to head with yours. Adjust your concentration until they're equally fast. There you have your units.
The method of converting fluorescence to amount of nucleotide incorporated is stated in the paper as follows: "...initial velocities from the fluorescence-based assay were calculated by using a standard curve for double-stranded DNA to estimated the amount (in ng) of double-stranded DNA formed per minute. Incorporation of deoxynucleotides (in pmol/min) was calculated by dividing the DNA amount by 2 (since only one of the two strands is synthesized by the polymerase) and using 324.5 Da as the average molecular weight of a deoxyribonucleoside monophosphate."
Adam B Shapiro : As you know fluorescence-based methods are semi-quantitative, is there a method that can quantify the dNTPs incorporated by Polymerase directly.
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