highQu dNTP sets and mixes meet all highest industry standards and allow for unrivaled performance of your PCR and other DNA synthesis, labeling or sequencing reactions.
Produced under the stringent quality monitoring conditions, they guaranty reproducible results. More than 99% HPLC purity eliminates inhibitions of PCR and allows for increased yields with higher dNTP concentrations.
Exceptional stability of dNTPs allows for short term ambient temperature shipments, short term room temperature storage or long PCR of more than 30 kb targets (see the picture of Long PCR obtained with highQu dNTPs) as well as long amplifications exceeding 40 cycles.
Preparation of dNTP Mixes from a Set of four 100 mM dNTPs:
- Highly concentrated solutions (like 100 mM dNTP Set) require thorough mixing before the use.
- The optimal dNTP mix shall have equal concentrations of all 4 dNTPs.
- To prepare from a set of 4 dNTPs desired mixes of common concentrations, follow the guidelines below:
|Use same volume of each from
four 100 mM dNTP solutions:
|Resulting 1 ml
|20 µl each||920 µl||2 mM dNTP mix|
|25 µl each||900 µl||2.5 mM dNTP mix|
|100 µl each||600 µl||10 mM dNTP mix|
|250 µl each||-||25 mM dNTP mix|
Protocol recommendations for standard PCR
- Typical concentration of each dNTP in the PCR reaction is 0.2 – 0.25 mM.
- Higher concentration increase yields, however Mg2+ ions bind to dNTPs, therefore, both components shall be present in coordinated concentrations.
- Too high dNTPs and magnesium concentrations reduce PCR fidelity.
- Mix well each dNTP and magnesium solution, to avoid concentration fluctuations.
- We recommend final 3 mM MgCl2 with 0.25 mM each dNTP concentration for routine PCR.
- Recommended volumes of dNTP Mixes and 50 mM MgCl2 are given below, however those are just guidelines, and each reaction might require optimization.
|Starting dNTP Mix conc.||Vol. of dNTP mix for 50 µl rxn to achieve 0.25 mM concentration of each dNTP|
|10 mM||1.25 µl|
|25 mM||0.5 µl|
|Final Mg2+ conc. in rxn||Vol. of 50 mM MgCl2 for 50 µl rxn to achieve desired conc.|
|2 mM||2 µl|
|3 mM||3 µl|
|NUS0101||0.25 ml - 100 mM dATP
0.25 ml - 100 mM dCTP
0.25 ml - 100 mM dGTP
0.25 ml - 100 mM dTTP
Aqueous solution of 100 mM dATP, pH 7.0.
In the dark at -20°C.
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