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Deoxyribonucleoside triphosphates (dNTPs)

  1. Dissolve each dNTP in distilled water at an approximate concentration of 100 mM.
  2. Adjust to pH 7.0 with 0.05 M Tris base using a micropipette (use pH paper to check the pH).
  3. 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

 

  1. 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.
  2. 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.

 

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