biotechrabbit™ deoxynucleotide triphosphates are first-choice nucleotides for all PCR applications, including the most demanding, such as amplification of long targets (up to 40 kb), GC-rich templates, qPCR, cDNA synthesis, high-fidelity PCR, DNA labeling and sequencing.
Advanced production technology ensures that deoxyribonucleotide triphosphates have >99% purity and outstanding stability, ensuring excellent performance and consistent, reliable results.
For the maximum flexibility, nucleotides are available in sets and mixes of common concentrations.
Component | Composition |
all dNTP Sets | Aqueous solution (pH 7.0) containing |
STORAGE | −20°C (until expiry date – see product label) |
Quality Control
Functional assay
Functionality tested in PCR.
Purity test
The concentration of each lot is verified by optical density spectrometry. The purity of nucleotide triphosphates is determined by HPLC. The minimum passing specification for the triphosphate content is 99%.
Contamination test
The absence of human and E. coli DNA is confirmed by qPCR.
Prevention of PCR contamination
When assembling the amplification reactions, care should be taken to eliminate the possibility of contamination with undesired DNA.
- Use separate clean areas for preparation of samples and reaction mixtures and for cycling.
- Wear fresh gloves. Use sterile tubes and pipette tips with aerosol filters for PCR setup.
- Use only water and reagents that are free of DNA and nucleases.
- With every PCR setup, perform a contamination control reaction that does not include template DNA.
Typical use of dNTPs
- 0.2 mM of each nucleotide is the recommended final dNTP concentration in common PCR applications.
- It is essential to have equal concentrations of all four dNTPs in the PCR mixture.
- Every time before the use thaw and mix well each dNTP solution, especially highly concentrated ones.
- To prepare on your own 1 ml dNTP mixtures of common 2 mM, 2.5 mM, 10 mM or 25 mM concentrations, follow the guidelines below:
1 ml Mix | 100 mM dATP | 100 mM dCTP | 100 mM dGTP | 100 mM dTTP | Water |
2 mM dNTP Mix: | 20 µl | 20 µl | 20 µl | 20 µl | 920 µl |
2.5 mM dNTP Mix: | 25 µl | 25 µl | 25 µl | 25 µl | 900 µl |
10 mM dNTP Mix: | 100 µl | 100 µl | 100 µl | 100 µl | 600 µl |
25 mM dNTP Mix: | 250 µl | 250 µl | 250 µl | 250 µl | - |
- Use the ready dNTP mixes in PCR (at 0.2 mM final concentration each) according to the guidelines below:
Volumes of dNTP mixes for the indicated PCR volume | ||||
dNTP Mix concentration | Used for PCR as | 50 µl PCR | 25 µl PCR | 20 µl PCR |
2 mM | 10× | 5 µl | 2.5 µl | 2 µl |
2.5 mM | 12.5× | 4 µl | 2 µl | 1.6 µl |
10 mM | 50× | 1 µl | 0.5 µl | 0.4 µl |
25 mM | 125× | 0.4 µl | 0.2 µl | 0.16 µl |
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