dNTP Mix, 100mM, Lithium Salt
Ready-to-use sequencing grade ultra-pure Deoxynucleotide triphosphate (dNTP) mix, 25 mM each of dATP, dCTP, dGTP and dTTP solution (100 mM total), supplied as lithium salts in purified water at pH 7.5, enzymatically synthesized from premium quality raw materials, using highly specific production systems in our purpose-built facilities. The manufacturing process eliminates impurities and PCR-specific inhibitors such as modified nucleotides, tetraphosphates, and pyrophosphates commonly observed in other commercially available dNTP products (>99% purity determined by quantitative HPLC). Lithium salts have greater resistance to repeated freezing and thawing cycles than sodium salts, and lithium salt dNTP preparations remain sterile over the entire shelf-life due to the bacteriostatic activity of lithium towards various microorganisms.
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Meridian's dNTP Mix, 100mM, Lithium Salt
- Greater than 99 % purity, ideal for use in qPCR applications
- Free from PCR inhibitors for maximum assay performance
- Enzymatically synthesized from premium quality raw materials
- Free from DNase, RNase and Nickases
Highly sensitive multiplex qPCR reaction
A 10-fold serial dilution of human genomic DNA amplified with four different probes, both in singleplex reactions (blue line) and quadruplex reaction (red line) using dATP, dCTP, dGTP and dTTP in lithium salt. The results illustrate the same high sensitivity, excellent reproducibility and Ct values for both the singleplex and multiplex reactions with no reduction of efficiency often associated with multiplexing.
dNTP Mix, 100mM (Lithium), MDX051
Ultra-pure mix of dATP, dGTP, dCTP, and dTTP in lithium salt solution, enzymatically synthesized from premium quality raw materials.
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Description
Ultra-pure dNTP (dATP (2’-Deoxyadenosine-5’-Triphosphate), dCTP (2’-Deoxycytidine-5’-Triphosphate), dGTP (2’-Deoxyguanosine-5’-Triphosphate) and dTTP (2’-Deoxythymidine-5’-Triphosphate)) lithium salt solution is enzymatically synthesized from premium quality raw materials, using highly specific production systems in our purpose-built facilities.
Specifications
| escription | Ultra-pure dNTP (dATP (2’-Deoxyadenosine-5’-Triphosphate), dCTP (2’-Deoxycytidine-5’-Triphosphate), dGTP (2’-Deoxyguanosine-5’-Triphosphate) and dTTP (2’-Deoxythymidine-5’-Triphosphate)) lithium salt solution is enzymatically synthesized from premium quality raw materials, using highly specific production systems in our purpose-built facilities. | |||
| dATP | dCTP | dGTP | dTTP | |
| Composition | C10H12N5O12P3Li4, | C10H12N3O13P3Li4 | C10H12N5O13P3Li4 | C10H13N2O14P3Li4 |
| Molecular weight | 514.916 g/mol | 490.891 g/mol | 530.916 g/mol | 505.903 g/mol |
| λmax (at pH 7.0) | 259 ± 1 nm | 272 ± 1 nm | 252 ± 1 nm | 267 ± 1 nm |
| A250/A260 | 0.78 ± 0.03 | 0.82 ± 0.03 | 1.16 ± 0.05 | 0.65 ± 0.03 |
| A280/A260 | 0.15 ± 0.02 | 0.98 ± 0.03 | 0.66 ± 0.03 | 0.73 ± 0.02 |
| Concentration
| 25 mM ± 5%
(at λmax, pH 7.0, ԑ = 15.4 E x mmol-1 x cm-1) | 25 mM ± 5%
(at λmax, pH 7.0, ԑ = 9.1 E x mmol-1 x cm-1) | 25 mM ± 5%
(at λmax, pH 7.0, ԑ = 13.7 E x mmol-1 x cm-1) | 25 mM ± 5%
(at λmax, pH 7.0, ԑ = 9.5 E x mmol-1 x cm-1) |
| Presentation | 1 vials | |||
| pH of Solution (at 20°C) | 7.5 – 8.0 | |||
| Appearance | Clear, colorless solution | |||
| Application | cDNA synthesis, PCR, long-range PCR (>20 kb), high fidelity PCR, RT-PCR, qPCR, RT-qPCR, LAMP, microarrays, long NGS | |||
| Consistency | Single band with PCR amplification of a 3 kb amplicon with a serial dilution of dNTP. | |||
| DNA contamination | None detected in qPCR amplification with traces overlay with the negative control on E. coli and mouse genomic DNA specific targets. | |||
| DNase/RNase/Nickase activity | None detected | |||
| Purity dNTP
(HPLC Area % at λmax) | ≥99% | |||
| dNDP + dNMP
(HPLC Area % at λmax) | <1% | |||
| DNase/RNase Contamination | No detectable degradation | |||
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FAQs: dNTP Mix, 100mM
Why is the purity of dNTPs important?
dNTPs or deoxynucleotide triphosphates are the “building blocks” for DNA. Purity and stability of dNTPs are two of the essential factors to achieve a successful PCR. The use of a highly purified dNTP preparation is particularly recommended for sensitive techniques such as long-range PCR, RT-PCR, multiplex PCR, mutagenesis experiments and real-time applications. The purity of dNTPs is also important when the starting amount of template is minimal.
What concentration of dNTPs is normally used in a PCR?
The standard concentration of a dNTP for PCR reactions is 0.2 mM. If the starting stock is a 25 mM solution of each dNTP in a 100mM dNTP Mix, you need to add 0.5 µL to a 50 µL standard PCR reaction.
Can I increase the DNA yield by adding higher concentrations of dNTPs?
It depends. You probably can increase the DNA yield, but you will have to optimize the complete PCR reaction, adjust the buffer, the Mg2+ and so on. It is not a matter pertaining only to nucleotides.
Does dNTP quality affect processivity?
Yes. The sodium salt dNTP products are a standard grade and so we recommend these dNTPs for PCR. For more sophisticated reactions such as amplification of long templates, real-time PCR and NGS, we recommend lithium salt dNTP products.
Is the pH of the dATP solution important for stability?
Yes. The optimal pH for storage of nucleotides is from pH 7.0-8.2 (pH at 20°C). An acidic pH will cause hydrolysis of dATPs (deoxyadenosine triphosphate) to dADPs (deoxyadenosine diphosphates) and dAMPs (deoxyadenosine monophosphates), rendering them less suitable for PCR applications. During freezing/thawing cycles, the pH of the dATP solution can differ from the pH at 20°C. Sodium salts are temperature sensitive, so care needs to be taken when repeatedly frozen and thawed and sodium salt dATP is better aliquoted into smaller volumes and kept frozen in order to extend their shelf life.
What are the advantages of enzymic synthesis over chemical synthesis of dNTPs?
The enzymatic synthesis of dNTPs uses highly specific enzymatic systems which eliminate impurities and PCR inhibitors, such as modified nucleotides, PPi and deoxynucleoside tetraphosphates. PCR reactions are impeded by the presence of contaminants resulting from chemical manufacturing processes, such as traces of dNDPs, pyrophosphates or other ionic species (e.g. acetate). Such contamination may lead to poor yields or to no PCR product at all. Unless thoroughly purified, chemically synthesized dNTPs often contain deoxynucleoside tetraphosphates which are powerful PCR inhibitors. Chemical synthesis can also lead to deamination and other nucleotide modifications whereas enzymatic synthesis of dNTPs bypasses these risks.
Are the concentrations of the dNTPs each or total?
The concentration of our dNTP Mix are total – 100 mM dNTP Mix is made up of 25 mM of each dNTP (dATP, dCTP, dGTP and dTTP).
I need to dilute my dNTPs to a different concentration, what should I use as a diluent?
We recommend that you dilute your dNTPs using molecular biology or PCR grade water.
Can your dNTP Mix be used for lyophilization?
Yes, all of our dNTPs are glycerol-free and so can be lyophilized.
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