Ideal for PCR, qPCR, RT-PCR, cDNA synthesis, NGS, isothermal amplification, hi-fidelity and long–range PCR.
Guide to Meridian Bioscience’s dNTPs Ultra-Pure Nucleotides
www.MeridianLifeScience.com
Contents Introduction Excellence in Development and Production Purity Performance Applications, Stability and Storage
Customized Solutions Ordering Information
Why Choose Meridian Bioscience’s dNTPs? • 99% trisphosphate purity determined by HPLC • Free from PCR inhibitors • DNase, RNase, Protease and Nickase free • Supplied as individual dNTPs, in sets and as mixes • Available in sodium or lithium salts • Extended shelf-life of 24 months at -20 o C • Custom, bulk and OEM nucleotides • Manufacturing site: Luckenwalde Biotechnology Park, Berlin-Brandenburg Life Science Cluster
As one of the few primary manufacturers of nucleotides in the world, Meridian brings over 30 years of expertise to assay development. Our ultra-pure dNTPs are synthesized from high-quality raw materials in state-of-the-art production facilities, ensuring ≥ 99% purity, as verified by quantitative high-performance liquid chromatography (HPLC). Stringent purification processes minimize cross-contamination, delivering the reliability and consistency essential for high-performance molecular assays. Manufactured under ISO 9001-certified quality standards, our nucleotides support robust and reproducible assay development at every stage.
ISO 9001:2008 Quality Management Certification Features • Validation Support Documentation • Rigorous QA/QC Systems
• Retention Samples • Product Traceability
• Customer On-Site Audits • Regular Inspections by Certifying Authorities
www.meridianbioscience.com/lifescience
Product Information
Quality Stringent control systems guarantee that Meridian’s dNTPs are of the highest quality and consistency to be found anywhere in the industry. A further assurance of this is the ISO certification of our purpose-built state-of-the-art nucleotide manufacturing facilities in the Luckenwalde Biotechnology Park, Berlin-Brandenburg Life Science Cluster, Germany. With ISO certification implemented, Meridian demonstrates its commitment to being a reliable and techni- cally competent partner, for our customers, suppliers and future collaborators. With increasing level of sophistication of molecular processes and regulatory requirements, it has become essential to achieve highest dNTP quality, with minimal batch-to-batch variations. Meridian’s ultra-high purity deoxynucleoside triphosphates (dNTPs) are manufactured to the highest purity standards and perfectly suited for a wide range of applications from PCR (including low copy, long range, qPCR), isothermal amplification, cDNA synthesis/ reverse transcription to next-generation sequencing (NGS). Purity Purity of dNTPs is an essential starting parameter for any highly sensitive diagnostic assays. Inhibitors and contaminants generally exert their effects through negative interaction with template, interference with enzymes such as DNA polymerases, or by other means, such as sequestration of essential co-factors. These can individually contribute to reducing the efficiency of the assay and cause it to fail overall. Meridian’s ultra-pure dNTPs are manufactured to the highest purity standards in the industry. Our dNTPs are enzymatically synthesized (Fig. 1) from premium quality raw materials, using highly specific production systems in our purpose-built facilities. The manufacturing process eliminates trace amounts of impurities and inhibitors that can affect performance. These include: • dNDPs (deoxynucleotide diphosphates) and dNMPs (deoxynucleotide monophosphates), that can act as competitive inhibitors of enzymes, leading to reduced extension efficiency (Fig. 2). • Modified dNTPs (e.g., oxidized or deaminated forms), leading to lead to base misincorporation and mutations. • Residual chemical contaminants from synthesis, these include protecting groups and organic solvents that can inhibit or denature enzyme activity and reduce efficiency.
• DNase and RNase contaminants that can affect cDNA synthesis (Fig. 3). • Nickases and proteases, which can compromise the template (Fig. 4).
Fig.1 Enzymatic synthesis of dNTP Monophosphorylated nucleotides (dNMPs) are converted to deoxynucleotide diphosphates (dNDPs) and then nucleoside triphosphates (dNTPs) using kinase proteins. The synthesized dNTPs can then be used directly in nucleic acid amplification.
dATP
dCTP
Chromatogram
Chromatogram
100
100
50
50
0
0
0 1
2 3 4 5 6 7 8 91011121314151617
0 1
234567891011
12 13 14 15 16 17
Minutes
Minutes
dGTP
dTTP
Chromatogram
Chromatogram
100
100
50
50
0
0
0 1
234567891011
12 13 14 15 16 17
0 1
234567891011
12 13 14 15 16 17
Minutes
Minutes
Fig.2 HPLC profile of ≥ 99% pure dNTPs HPLC analysis shows greater than 99% triphosphate purity with undetectable mono-, di- and triphosphate forms.
Fig. 3 Functional purity of Meridian ultra-pure dNTPs in RT-PCR A ten-fold serial dilution of mouse total RNA (30ng to 3pg) was reverse transcribed using MMLV-RT (MDX044) and a1 kb fragment of RN18S amplified using Taq HS DNA Polymerase (MDX008) and run on a 1% agarose gel, showing the sensitivity of the assay using Meridian dNTPs
Amplication Plot
1,200,000 1,100,000 1,000,000
900,000 800,000 700,000 600,000 500,000 400,000 300,000 200,000 100,000 0 2 4 6 8 10 12 13 16 1820222426283032343638404244 Cycle
Fig. 4 Exceptional performance of Meridian ultra-pure dNTPs in quantitative PCR (qPCR) Activity of the Air-Dryable 1-Step RT-qPCR Mix (MDX095) in both wet (blue) and air-dried (red) formats were compared on a ten-fold dilution of mouse RNA. The air-dried mix showed no loss of activity and sensitivity when compared to freshly prepared wet mix up to the assay limit of detection. The high quality of dNTPs helps ensure consistent Ct values, especially with low template concentrations.
www.meridianbioscience.com/lifescience
Product Specification Table dATP
dCTP
dGTP
dTTP
dUTP
dATP lithium 100mM solution 2’-deoxyadenosine – 5’-triphosphate
dCTP lithium 100mM solution 2’-deoxycytidine – 5’-triphosphate
dGTP lithium 100mM solution
dTTP lithium 100mM solution dTTP lithium 100mM solution
dUTP lithium 100mM solution dTTP lithium 100mM solution
Product
2’-deoxyguanosine-5’- triphosphate
Nomenclature
C 10 H 12 N 5 O 12 P 3 *
C 9 H 12 N 3 O 13 P 3 * 490.9g/mol 100mM ± 5%
C 10 H 12 N 5 O 13 P 3 * 530.9g/mol 100mM ± 5%
C 10 H 13 N 2 O 14 P 3 * 505.9g/mol 100mM ± 5%
C 9 H 12 N 2 O 14 P 3 * 492.884g/mol 100mM ± 5%
Formula
Molecular Weight
514.9g/mol
Concentration Appearance pH of Solution Purity (HPLC)
100mM ± 5%
Clear Colorless Solution Clear Colorless Solution Clear Colorless Solution Clear Colorless Solution Clear Colorless Solution
7.5
7.5
7.5
7.5
7.5
≥ 99%
≥ 99%
≥ 99%
≥ 99%
≥ 99%
dNDP (HPLC Area)
<1%
<1%
<1%
DNase, RNase λ max pH 7.0
Negative
Negative
Negative
Negative
Negative
259nm
272nm
252nm
267nm
262nm
15.4 E x mmol -1 x cm -1
9.1 E x mmol -1 x cm -1
13.7 E x mmol -1 x cm -1
9.6 E x mmol -1 x cm -1
10.0 E x mmol -1 x cm -1
ε at λ max @ pH7.0
A 250 /A 260 A 280 /A 260 Storage Stability
0.78 ± 0.03 0.15 ± 0.02
0.82 ± 0.03 0.98 ± 0.02
1.16 ± 0.05 0.66 ± 0.02
0.65 ± 0.03 0.73 ± 0.02
0.75 ± 0.03 0.38 ± 0.02
at -20°C
at -20°C
at -20°C
at -20°C
at -20°C
≤ 24 months
≤ 24 months
≤ 24 months
≤ 24 months
≤ 24 months
*All dNTP formulations are designed for convenience and flexibility. Individual nucleotides (dATP, dCTP, dGTP, dTTP and dUTP) are supplied in lithium (Li4) or sodium (Na3) salts. Applications Meridian ultra-pure dNTPs possess the highest level of purity in the industry and have been extensively validated for use in a wide variety of molecular biology applications. Validated applications PCR, RT-PCR qPCR, RT-qPCR Long-range & High-fidelity PCR cDNA synthesis Isothermal amplification Cloning Next-generation sequencing Sanger sequencing Genotyping Stability and Storage
A
Batch 1
Batch 2
Meridian ultra-pure dNTPs are available in sodium or lithium salts in highly purified water at pH 7.5 to ensure functionality and long-term stability. Our QC results demonstrate that Meridian’s dNTP functionality is not only retained after multiple freeze-thaw cycles but can also greatly exceed the standard 2-year storage recommendation time in a –20°C constant-temperature freezer.
5 cycles
B
10 cycles
Year 0
15 cycles
Year 2
20 cycles
Fig. 5 Meridian’s dNTP mix remains stable after multiple rounds of freeze-thawing and long-term storage. A) Two different batches of Meridian 100 mM dNTP mix were freeze-thawed up to 20 times and a two-fold serial dilution was used in a PCR reaction with 10 ng of λ DNA and Taq DNA polymerase to amplify a 2 kb fragment. Reactions with both batches produced identical PCR products, whether freeze-thawed once or multiple times. B) Stability of dNTPs were also assayed for PCR functionality for dNTPs stored at -20oC for up to two years. All dNTP batches performed exceptionally in PCR reactions with λ DNA (10 ng) and amplified with Taq DNA polymerase regardless of age of the dNTPs.
Customized dNTP Solutions When your requirements are beyond the scope of our standard product range we invite you to take advantage of our proven world-class bulk, custom and OEM service. Our aim is to provide the best customer service and dNTPs products in the industry. Being a primary manufacturer of nucleotides, we can accommodate requests for micro-liter to multi-liter quantities. We can manufacture special nucleotide formulations, blends and mixes to your requirements. Private labelling and packaging arrangements are also available. Manufactured under ISO 9001:2008 regulations, Meridian ultra-pure dNTPs cost-effectively bring you the quality and supply security to meet your needs. Flexibility at your service Scale • From micro-liters to multi-liters • From mgs to kgs • Special synthesis and batches Format • Any concentration • Any combination, formulation • Any volume • Custom mixes and blends Production Planning • Batch reservation • Lot retention samples dATP dCTP
• Special testing as required • Scheduled & global delivery Packaging and Labeling • Any size • Choice of tubes, vials & bottles • Custom labeling • Label design and printing
dGTP
dTTP
• Customized packaging • OEM product finishing Service and Support • Validation support file • Customer on-site audits • Product traceability • Confidentiality
www.meridianbioscience.com/lifescience
Ordering information Product
Conc.
Cat No
Individual dATP (lithium salt) dCTP (lithium salt) dGTP (lithium salt) dTTP (lithium salt) dUTP (lithium salt) dATP (sodium salt) dCTP (sodium salt) dGTP (sodium salt) dTTP (sodium salt) dUTP (sodium salt)
100 mM 100 mM 100 mM 100 mM 100 mM 100 mM 100 mM 100 mM 100 mM 100 mM
MDX046 MDX047 MDX048 MDX049 MDX059 MDX063 MDX064 MDX065 MDX066 MDX096
Set (dATP, dCTP, dCTP, dTTP) dNTP Set (lithium salt)
100 mM total
MDX050
Mix dNTP Mix (lithium salt) dNTP Mix (lithium salt) dUTP Mix (lithium salt) dNTP Mix (sodium salt) dNTP Mix (sodium salt) dNTP Mix (sodium salt)
10 mM total 40 mM total 50 mM total 10 mM total 40 mM total 100 mM total
MDX051 MDX085 MDX058 MDX067 MDX083 MDX084
For more information about Meridian ultra-pure nucleotides please visit: https://www.meridianbioscience.com/lifescience/products/molecular-reagents/nucleotides
Technical Support If you would like any support, help or technical advice we would like to like hear from you: UK: +44 (0)20 8830 5300 US: +1 901 382 8716 DE: +49 (0) )337 160222 00 or email us at: mbi.tech@meridianlifescience.com For bulk and OEM service enquiries please contact: mbi.tech@meridianlifescience.com
Ordering information: USA 5171 Wilfong Road Memphis, Tennessee 38134 Phone: +1 901-382-8716 Fax: +1 901-333-8223
Email: info@meridianlifescience.com Orders: orders@meridianlifescience.com www.meridianbioscience.com/lifescience
Connect with us:
ISO 13485 Certified
05/25
Page 1 Page 2 Page 3 Page 4 Page 5 Page 6 Page 7 Page 8Powered by FlippingBook