Original manufacturer of quality dNTP's
High Performance Nucleotides
› Primary manufacturer
› Custom formulations
› Micro to multi-litre solutions
› Exceptional performance
Because nucleotides don’t get better than this.
› Ultrapure
› Exceptional performance
› Stable for 2 years at -20°C
› Custom Formulations
Specifications
dNTP Solutions
Production Technology
Jena Bioscience is one of only a few primary manufacturers of dNTPs for PCR. Our high quality starts with our production technology. Many problematic impurities, such as pyrophosphate and modified nucleotides, are by-products from chemical synthesis. These impurities can severely impact PCR performance. That’s why we synthesise all our dNTPs enzymatically, meaning many common impurities are never even present in our solutions.
Any remaining impurities are removed with several state-of-theart purification procedures. For dATP, dCTP, dGTP, and dUTP, we start with the respective ribonucleotide, and use the highly specific Ribonucleotide Reductase enzyme (Scheme 1). While for dTTP, we use enzymes to sequentially phosphorylate thymidine.
Quality
The level of sophistication in PCR applications constantly reaches new highs. Performance can be negatively affected by even one poor quality reagent. That’s why we ensure the highest quality in our dNTP solutions.
According to our ISO 9001:2015 certified quality management system each dNTP lot is assayed under stringent criteria for purity and functionality (see Specifications, page 6). The quality of a dNTP solution can be assessed with three criteria: Purity, Macromolecular Contaminants, and Inorganic Species.
Purity
Purity of dNTPs is a deciding factor in PCR performance. A specification of ≥ 99.0 % dNTP is the market standard, yet this only tells you half the story. The constitution of the remaining ≤ 1.0 % is crucial. Common contaminants in dNTP solutions are:
§ Deoxyribonucleotide diphosphates – dNDPs
§ Ribonucleotides – NTPs
§ Other dNTPs – e.g. deaminated or methylated dNTPs § Other nucleosidic compounds
§ Phosphates Of these contaminants, only the dNDPs minimally affect PCR performance. Even trace quantities of other impurities can significantly affect performance
Macromolecular Contaminants
The absence of human DNA and bacterial DNA in dNTP solutions is critical. These macromolecules can be present from the use of bacterial enzymes used in production, as well as human DNA from handling during production.
Since the presence of just a few copies of genomic DNA can cause false positives in PCR, we analyse each dNTP lot at the end of production to ensure they are DNA-free. (Figure 1) In addition, we test for residual enzymatic activity to ensure absence of DNases, RNases, Nicking activities, as well as proteases.
Inorganic Species
The presence of critical concentrations of inorganic species may result from contaminated raw materials and inadequate manufacturing processes. These species commonly interfere with PCR and are commonly referred to as “PCR inhibitors”.
These do not absorb UV light and are not detected by UV-detection in reverse phase HPLC. To ensure our products are free from inhibiting quantites of these species, we use Analytical Anion Chromatography, GC/FID, and ICP-MS for analyses.
Supplier Comparison
To ensure our position as market leaders in quality, we routinely carry out extensive comparisons with other dNTP suppliers. These tests cover both purity and functionality.
Purity
To accurately assess purity, we have optimised HPLC methodology to allow for separation of similar components. Figure 2 shows a example comparison of Jena Bioscience dATP solution to four dATP solutions from other suppliers. The peaks for dATP and dADP are labelled; other peaks present are potentially harmful to PCR performance.
Due to the complexity of dNTP impurities, it is important to better illustrate purity. We first compared all four dNTPs (dATP, dCTP, dGTP, and dTTP) for each supplier. These values were then used to show the constitution of a dNTP mix for each supplier (Figure 3).
Summary
- All suppliers have a dNTP purity over 99.5 %
- Jena Bioscience has a higher quantity of dNDP – essentially harmless to PCR
- Jena Bioscience has a significantly lower quantity of other impurities – those reducing PCR performance
Functionality
Functionality and performance in PCR assays is the most important factor for dNTP solutions. The four dNTPs (dATP, dCTP, dGTP, and dTTP) from each supplier were used to create a dNTP mix (25 mM each). These mixes were first tested in an 18 kb PCR reaction - amplification of such a large DNA fragment requires exceptional performance and can be negatively affected by impurities in dNTP solutions (Figures 4).
Nucleotide impurities, such as NTPs, and modified dNTPs can strongly inhibit PCR enzymes. Therefore we use an assay particular sensitive to nucleotide impurities to evaluate all our dNTP solutions. The comparison of the dNTP mixes in this assay are shown in Figure 5.
Summary
- Jena Bioscience dNTP solutions gave clear bands in the 18 kb amplification.
- Jena Bioscience dNTP solutions performed exceptionally well in the assay sensitive to nucleotide impurities.
Test the exceptional performance of our nucleotides for yourself.
Request a sample!
Our entire product range is available as samples. We can provide multiple production lots of the following:
› Individual dNTP solutions (100 mM)
› dNTP mixes (25 mM each or 10 mM each)
› NTP solutions (100 mM)
› Custom formulations
› Lyophilisates
Contact us today at Sales@2BScientific.com