Contains 0.25 ml of 100 mM ultrapure dUTP. dUTP can either be fully substituted for dTTP or added in addition to conventional dNTPs. dUTP can be used with Family A polymerases (such as Taq) or modified Family B polymerases, such as Q5U Hot Start High-Fidelity DNA Polymerase. Q5U contains a mutation in the uracil-binding pocket that enables the ability to read and amplify templates containing uracil and inosine bases.
For a simplified protocol, dUTP can be added to most polymerase reactions at the same concentration as other nucleotides (e.g., a 1:1:1:1:1 dA/dC/dG/dT/dU ratio).
For applications that require other molar ratios (up to and including full substitution of dU for dT), custom nucleotide mixes must be made. We recommend starting with a concentration of 0.2 mM of each dNTP for both Taq and Q5U DNA polymerases.
Ultra purified water as a sodium salt at pH 7.5.
The dUTP Solution is supplied at a concentration of 100 mM (+/- 5 mM) as determined by A260 absorbance.
Can be diluted using sterile distilled water, preferably Milli-Q® water or can be diluted using sterile TE [10 mM Tris-HCl, 1 mM EDTA (pH 7.5)].
The following reagents are supplied with this product:
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