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Trouble-shooting: Low yield

50 40-45 Back to Top Nucleotides hydrolized

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Always store nucleotide stock solutions at a concentration of at least 10 mM, 100 mM is best. Significant hydrolysis occurs after storing at 1 mM for 2 months. Dissolve NTPs or dNTPs in water at an expected concentration of 10 mM. Using 0.05 M Tris-base and pH paper adjust the pH to 7.0. Dilute an aliquote of the neutralized NTP or dNTP appropriately and read the optical density at the wave lengths given in the following table. Calculate the actual concentration using the values for the extinction coefficients. Freeze away in small aliquots at -20C.

Base
Wavelength Extinction coefficients for bases
e(M-1 cm-1)
A 259 1.54 x 104 T 260 7.4 x 103 G 253 1.37 x 104 C 271 9.1 x 103 U 262 1.0 x 104

The lithium and sodium salts have equivalent st
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