DNA Quantification

From OpenWetWare

Revision as of 17:27, 6 April 2010 by Michael A. Speer (Talk | contribs)
Jump to: navigation, search
back to protocols



This protocol uses a spectrophotometer to quantify the amount (μg/mL) of DNA and then uses a simple equation to convert this mass concentration into a molar concentration. The molar concentration is much more useful for most enzymatic processes.

  • Example: digesting 500ng of a 2KB plasmid is twice as much "Enzymatic work" as digesting 500ng of a 4KB plasmid with the same multiple cloning site.


1. Dilute the DNA sample 30X by combining the following in a cuvette:

  • 87µl water
  • 3µl DNA prep

2. Run the DNA quantification (260/280) test on the spectrophotometer with a dilution of 30.
3. Make sure that the A260 measurement is between 0.1 and 1.
4. If is too low then repeat the measurement using 15X dilution.

  • 84µl water
  • 6µl DNA prep

5. Calculate the molar concentration of DNA using the following equation:

  • Picomoles/µl = DNA Concentration(µg/ml) / [0.66*DNA Size(bp)]



Personal tools