Meridian is a primary manufacturer of specialized high-quality molecular reagents and offers solutions to a wide range of industries to diagnose and treat diseases, discover new therapeutics or develop tests for environmental, food and cosmetic safety.
High-performing DNA polymerases
Selecting the right enzyme is critical to the success of a molecular diagnostic assay. At Meridian, our team of scientists are dedicated to creating cutting-edge enzymes that maximize efficiency, reliability and accuracy. With a keen focus on precision and performance, we continuously refine our techniques to deliver reagents that exceed expectations in every aspect. Choosing the right DNA polymerase DNA polymerases are used for DNA amplification in PCR (polymerase chain reaction) assays supporting applications such as sequencing, genotyping, pathogen detection, and quantitation. There are many different types of DNA polymerases, each with different properties and characteristics including thermal stability, specificity, processivity, fidelity, proofreading and elongation rate. They are derived from various organisms, such as bacteria, archaea, eukayotic cells and viruses, evolving over billions of years to perform specialized functions unique to the organism they are derived from. Depending on the type of molecular assay, certain DNA polymerase characteristics are sought after, while others may be less desirable, which is why selecting the right enzyme is essential to achieving the correct assay performance.
MDX001
MDX008
MDX011
MDX015 MDX003 MDX203 MDX009 MDX012
MDX018
Template Length
5kb
5kb
5kb
5kb
5kb
2.5kb
5kb
5kb
5kb
Antibody (Seperate)
Hot Start
No
Antibody
Aptamer
Antibody
No
Chemical
No
No
1x
1x
1x
1x
9x
9x
1x
1x
1x
Fidelity
Processivity
High
High
High
High
High
High
High
Concentration
5 U/ µ L
5 U/ µ L
50 U/ µ L 50 U/ µ L
2 U/ µ L
20 U/ µ L
5 U/ µ L
5 U/ µ L
5 U/ µ L
Glycerol
50%
50%
0%
0%
50%
0%
50%
50%
0%
Strand displacement Low bioburden Routine PCR qPCR Isothermal Amplification High Specificity Multiplex GC-rich High-Fidelity NGS library amplification Lyophilization / Air drying
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PCR & qPCR Applications
Taq DNA Polymerase & Taq Hot-Start (HS) DNA Polymerase
Taq DNA polymerase is from the bacterium Thermus aquaticus and is a heat-stable enzyme commonly used in PCR. Meridian’s Taq DNA polymerase is designed for fast and sensitive PCR from complex templates and is supplied with a novel buffer containing dNTPs, MgCl 2 and enhancers that have been optimized for robust inhibitor tolerance and fast cycling conditions, considerably reducing the reaction time without compromising on PCR sensitivity or yield. It is also available as an antibody-mediated hot start enzyme, ensuring a reaction remains completely inactive during PCR set-up to prevent non-specific amplification.
> Fast DNA polymerase, delivering high yield under very fast PCR cycling conditions (less than 30 min) > Novel optimized buffer system maximizes the efficiency of PCR amplification, even in the presence of PCR inhibitors > Ideal for amplifying any target up to 5 kb, including DNA extracted from human, animal and plant samples
Figure 1. Taq DNA Polymerase: Robust amplification of GC-rich DNA
Figure 2. Taq DNA HS Polymerase: Sensitive amplification across a 10-fold serial dilution
A serial dilution in duplicate of human genomic DNA (1 μ g-12.5 ng, 1-6 respectively) was used with Taq DNA Polymerase to amplify 61% GC rich fragment human myc gene, to demonstrate the high yield and sensitivity of the Taq DNA Polymerase.
10-fold serial dilution (10 4 copies down to 10 copies) of human DNA using Taq HS DNA Polymerase and Fast qPCR Buffer, 4x (MDX033) to illustrate the speed, sensitivity and reproducibility when using Taq HS DNA Polymerase in a qPCR reaction.
PRODUCT
CAT NO.
VOLUME
REACTIONS
0.1 mL
500 Rxns
MDX001
Taq DNA Polymerase
10 mL
50,000 Rxns
0.1 mL
500 Rxns
MDX008
Taq HS DNA Polymerase
10 mL
50,000 Rxns
Glycerol-Free Taq Hot-Start DNA Polymerase
Meridian’s Glycerol-Free Taq DNA polymerase is a high-concentration (50 U/ µ L) enzyme optimized to deliver robust amplification without glycerol. It is supplied as a separate 10x glycerol‑free Taq DNA Polymerase, glycerol‑free Taq antibody, and glycerol‑free enzyme dilution buffer, making it compatible with lyophilization for assay manufacturers developing room-temperature stable assays. The high enzyme concentration allows for bulk production of PCR master mixes and the glycerol-free format is favorable when using automated pipetting robots and other applications where accurate pipetting of small volumes is crucial. > High concentration (50 U/ µ L) enzyme supplied as a separate Taq, hot-start antibody, and enzyme dilution buffer, all glycerol-free, for the ultimate flexibility in assay development > Ideal for efficient large-scale production of PCR master mixes and applications using automated pipetting robots or where accurate pipetting of small volumes is critical > Compatible with lyophilization for developing ambient-temperature stable assays
Figure 3. Glyercol-Free Hot Start Polymerase: Efficient and sensitive amplification used as wet mix or after lyophilization
Figure 4. Enzyme maintains stability after freeze-thaw cycles
110
100
90
80
70
60
50
0
10
15
F/T Cycles
Glycerol is a cryoprotectant and is normally part of the storage buffer, where it serves to protect Taq polymerase during freezing conditions. To test the resistance of Glycerol‑Free Taq Hot‑Start to freezing and thawing, a freeze‑thaw test was performed (10 and 15 cycles of freeze/thawing) and compared to fresh product (0 cycles of freeze/thawing). The results illustrate that Glycerol‑Free Taq Hot‑Start offers the same protection to freeze/thawing as a polymerase that contains glycerol.
A 10-fold serial dilution of human DNA (10 3 copies down to 10 copies) using Glycerol-Free Taq HS 50U/ μ L as a wet mix ( black ) or lyophilized mix ( blue ) to illustrate that lyophilization does not have an effect on the quality of the Glycerol-Free Taq HS.
PRODUCT
CAT NO.
VOLUME
REACTIONS
0.02 mL
1,000 Rxns
Glycerol-Free Taq HS, 50 U/ µ L
MDX011
5 mL
25,000 Rxns
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PCR & qPCR Applications
Aptamer Taq Hot-Start Polymerase (Glycerol-Free)
A high concentration (50 U/ µ L) glycerol‑free Taq DNA Polymerase with a DNA aptamer that serves as a molecular switch to regulate the polymerase’s activity. Unlike traditional hot start methods that require activation for several minutes at very high temperatures, aptamers can immediately disassociate from the polymerase (e.g. activate it) at relatively low temperatures (45 °C). The aptamer reversibly binds to the polymerase and inhibits its activity at room temperature before and after reaction completion, providing tighter control over the start of the PCR reaction and enhancing its efficiency and specificity. Aptamer Taq HS is an ideal polymerase for challenging high-throughput assays where fast reaction protocols and high specificity are required.
> Uses aptamer technology acts as a molecular switch to regulate the polymerase’s activity and reduces assay run time by up to 15 minutes > Convenient room-temperature reaction set-up > Highly suited to multiplex, high-throughput viral detection assays requiring high specificity
Figure 5. Aptamer Taq: Specific, sensitive and reproducible amplification in a multiplex assay
Figure 6. Aptamer Taq: Fast Hot-Start
Four viral sequences, two DNA (Cytomegalovirus ( green line ) and Adenovirus ( blue line )) and two RNA (Rotavirus ( orange line ) and Norovirus ( red line )) were amplified using Glycerol Free Instant Taq HS (MDX015) using Lyo-Ready qPCR Buffer (MDX022) and RNase-Tolerant MMLV-RT (MDX043) in quadruplex qPCR probe assays. The results illustrate that the use of aptamer hot-start gives just as sensitive results with RNA as it does with DNA templates.
Comparison of qPCR performance of Aptamer Taq HS (Glycerol-Free) ( dark blue ) and an antibody hot-start Taq (MDX008) light blue ). qPCR reactions were run with and without an initial 2-minute, high-temperature activation step for both polymerases and Ct values were compared. The data illustrates the immediate activation of the Aptamer Taq HS (Glycerol-Free), with no differences in Ct values with and without an activation step, unlike the antibody hot-start Taq, allowing for faster hot-start and faster reaction protocols without compromising sensitivity.
PRODUCT
CAT NO.
VOLUME
REACTIONS
0.1 mL
500 Rxns
Aptamer Taq HS (Glycerol-Free)
MDX015
10 mL
50,000 Rxns
Low DNA Taq HS 5 U/ µ L
A highly purified, chemically modified hot-start Taq DNA polymerase. Ideal for room-temperature PCR set-up and assays requiring low bioburden (i.e. assays for bacterial targets). Supplied in a reaction buffer containing dNTPs and enhancers, with MgCl 2 as a separate component for easy optimization. The low DNA background and stringent hot-start properties of Low DNA Taq HS are ideal for applications that are susceptible to false positives such as water testing and microbial testing. > Chemical hot-start Taq polymerase, ideal for room-temperature assay set-up minimizing primer dimerization or mispriming > Highly-suited for microbial tests that require low-burden reagents > Ideal for low-copy target amplification (such water testing)
Figure 7. Low DNA Taq: Highly sensitive and specific amplification.
A 10-fold dilution of DNA (10 3 copies down to 10 copies) was used in a qPCR assay using Low DNA Taq HS and an intercalating dye. Each dilution was run in triplicate using standard reaction conditions. A/ The amplification curve demonstrates the reproducibility and sensitivity of the Low DNA Taq HS and B/ The single distinct peak in the melt curve illustrates the products are a single discrete species, with no additional bands or primer/dimers.
PRODUCT
CAT NO.
VOLUME
REACTIONS
0.1 mL
500 Rxns
MDX009
Low DNA Taq HS 5 U/ µ L
10 mL
50,000 Rxns
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PCR & qPCR Applications
High-Fidelity Pfu
A high-fidelity antibody-mediated hot-start DNA polymerase from the bacteria Pyrococcus furiosus (Pfu). The Pfu polymerase enzyme replicates DNA in the 5´ → 3´ direction and also possesses 3´ → 5´ exonuclease (proofreading) activity. Base misincorporations that may occur during polymerization are rapidly excised by this proofreading activity. Meridian’s High-Fidelity has an error rate of 3.0 x 10 -6 generating blunt-ended amplicons up to 5 kb in length. It is supplied in a novel buffer system containing dNTPs and enhancers with MgCl 2 provided separately to enable flexibility for applications such as target enrichment, NGS library amplification, or cloning.
Figure 8. High-Fidelity Pfu: NGS library amplification using E.coli DNA
> Antibody hot-start Pfu polymerase for increased specificity > Error rate of 3.0 x 10 -6 > Ideal for NGS library amplification
PRODUCT
CAT NO.
VOLUME REACTIONS
Four NGS libraries were created using E. coli DNA. The libraries were amplified using High-Fidelity Pfu (MDX003) prior to being placed on flow cells and sequenced. For each sample, the fraction of uniquely mapped (mapped to only one place on a reference sequence), ambiguously mapped and unmapped reads relative to the total number of reads per sample is shown. The results illustrate that with E. coli DNA, High-Fidelity Pfu allowed for over 90% unique reads that could be aligned to the reference sequence.
0.2 mL 500 Rxns
MDX003
High-Fidelity Pfu
10 mL 25,000 Rxns
Glycerol-Free High-Fidelity Pfu
A high‑concentration (20 U/ µ L), glycerol‑free, high-fidelity thermostable Pfu enzyme supplied in a novel buffer system containing magnesium, dNTPs and PCR excipients. With an error rate of 3.0 x 10 -6 , this enzyme is ideal for enhancing DNA target enrichment in NGS library fragments and minimizing the incorporation of mutations into the NGS library. Combined, these characteristics facilitate the need for less extensive sequence coverage, leading to notable savings in both time and costs.
Figure 9. Glycerol-Free High-Fidelity Pfu: Efficient multiplex amplification used as wet mix or after lyophilization
> High concentration (20 U/ µ L) Pfu polymerase allowing for bulk production of PCR master mixes > Ideal for NGS library amplification > Compatible with lyophilization for developing ambient-temperature stable assays
PRODUCT
CAT NO.
VOLUME REACTIONS
0.1 mL 500 Rxns
Glycerol-Free High- Fidelity Pfu (HC)
MDX203
Glycerol-Free High-Fidelity Pfu (HC, MDX203), was lyophilized with Lyo-Ready ™ Pfu Reaction Buffer, 5x and stored at 37°C for 7 and 28 days. Lyophilized samples were tested in a multiplex PCR reaction amplifying 7 targets (793 bp, 649 bp, 548 bp, 418 bp, 332 bp, 196 bp, 135 bp) against the same enzyme and reaction buffer without lyophilization (Wet Reference) to demonstrate the enzyme’s stability after lyophilization.
10 mL 50,000 Rxns
Isothermal Applications
Bst DNA Polymerase
Bst DNA Polymerase, short for Bacillus stearothermophilus DNA Polymerase, is a type of enzyme commonly used in isothermal DNA amplification methods such as Loop-Mediated Isothermal Amplification (LAMP), Isothermal Helicase-Dependent Amplification (HAD) and Isothermal Multiple Cross Displacement Amplification (MCA). Meridian’ Bst DNA Polymerase is derived from the large fragment of Bacillus stearothermophilus DNA Polymerase I and exhibits fast 5´- 3´ DNA polymerase activity and strong strand displacement activity to deliver the fastest amplification speed (time-to-results), yield, salt tolerance, and sensitivity. It is available in two concentrations, 8 U/ µ L or 100 U/ µ L and is supplied with a separate Enzyme Dilution Buffer and a Bst Reaction Buffer for complete assay design flexibility.
> Optimized for isothermal amplification such as LAMP, HAD and MCA > Designed for fast polymerization and quicker time-to-results > Supplied with a separate reaction buffer and enzyme dilution buffer > Available in two concentrations, including a high 100 U/ µ L concentration
Figure 10. Bst DNA Polymerase: Faster time-to-results and better reproducibility compared to competitors
Figure 11. High-concentration Bst DNA Polymerase: Faster time-to-results and better reproducibility compared to competitors
CFTR (cystic fibrosis transmembrane receptor) was screened using High Conc. Glycerol-Free Bst ( orange ), NEB Bst 2.0 ( purple ), NEB wild-type Bst Large Fragment ( green ) and ArcticZymes IsoPol DNA Polymerase ( blue ), using the same test conditions across four technical replicates, to demonstrate the faster time to results (TTR) and better reproducibility of High Conc. Glycerol-Free Bst (MDX018).
A human pathogenic parasite (Strongyloides stercoralis) was screened using Bst Polymerase ( orange ), NEB Bst 2.0 ( purple ) and NEB wild-type Bst Large Fragment ( green ), using the same test conditions across four technical replicates, to demonstrate the faster time to results (TTR) and better reproducibility of Bst Polymerase (MDX012).
PRODUCT
CAT NO.
VOLUME
REACTIONS
1 mL
8,000 Rxns
MDX012
Bst DNA Polymerase
10 mL
80,000 Rxns
0.08 mL
8,000 Rxns
High Conc. Glycerol-Free Bst (100 U/ µ L)
MDX018
0.8 mL
80,000 Rxns
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
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08/23
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