- Dissolve each dNTP in distilled water at an approximate concentration of 100 mM.
- Adjust to pH 7.0 with 0.05 M Tris base using a micropipette (use pH paper to check the pH).
- Dilute an aliquot of the neutralized dNTP appropriately, and read the optical density at the wavelengths given in the table below.
Base Wavelength Extinction coefficient (M-1CM-1)
A 259 1.54×104
G 253 1.37×104
C 271 9.1×103
T 260 7.4×103
- Calculate the actual concentration of each dNTP. For a cuvette with a path length of 1 cm, absorbance is equal to the product of the extinction coefficient and the molar concentration.
- Dilute the solutions with distilled water to a final concentration of 50 mM dNTP.
<STORAGE> Store each separately at -70 C in aliquots of appropriate volume.