Protocol

Preparation of deoxynucleotide triphosphate solutions

The concentration of the ultrapure dNTPs is only approximately 100 mM.

1. Thaw the 100 mM dNTP completely
2. Make a stock solution of the dNTP by mixing 50 mL dNTP and 950 mL H₂O. The concentration will nominally be 5 mM.
3. Add 10mL of this solution to 990 mL of either H₂O or 0.1 M HCl is measuring cytosine and its derivatives.
4. Measure the absorbance spectra from 230 to 340 nm and determine the absorbance at the appropriate wavelength according to Table 1.
5. The concentration of the solution can be derived from Beers Law: A = ecl, in which A is the absorption, e the molar extinction coefficient, c the concentration, and l the pathlength.