Master Mixes for Molecular Ambient-Temperature Stable Assays
DNA Polymerases and qPCR/RT-qPCR Master Mixes Formulated for Lyophilization or Oven/Air-Drying
KEYAPPLICATIONS
Human Diagnostics
Blood Banking
Genetic Screening
Vet Health
Environmental
Water Testing
Food Testing
Molecular diagnostic tests are progressively moving towards lyophilized and air-dried formats. There are several advantages for this, including room temperature shipping and storage, extended shelf-life and increased flexibility in sample volume. In order to be compatible with drying however, enzyme preparations must be glycerol-free and include specialized excipients that preserve the mixture as it is exposed to various conditions including freezing, temperature ramps, vacuum
and dehydration. An ideal lyophilization formulation for example, should stabilize an enzyme in a freeze-dried format and allow very fast rehydration and reactivation of the enzyme preparations, without impacting its performance post rehydration.
Blood
Saliva
Urine
Stool
www.meridianlifescience.com
Air-Dryable
PRODUCT FEATURES:
• Glycerol-free mixes containing Taq polymerase, reaction buffer, dNTPs, MgCl 2 , and air-dry excipients • Ideal for developing extraction-free assays for direct detection from the clinical sample • Suitable for multiplex assays on POC diagnostic platforms or automated high-throughput instruments • Mixes can be used as a liquid or air-dried to create ambient-temperature stable assays • Compatible with a range of air-drying protocols • Simple, easy to use, and less costly than other forms of drying
Air-Dryable ™ qPCR Mix | Applications:
• Inhibitor-tolerant mixes designed for direct amplification from a wide range of specimens • Specimens require minimal sample processing, no extraction required PRODUCT HIGHLIGHTS Full enzyme activity following air-drying Activity of Air-Dryable qPCR Mix in both liquid and air-dried formats were compared by singleplex qPCR assay on 10-fold dilution mouse cDNA template. The air-dried mix showed no loss of activity and sensitivity when compared to freshly prepared liquid mix up to the assay limit of detection.
PRODUCT
CAT NO. VOLUME REACTIONS
5 mL
1,000 Rxns
Air-Dryable ™ qPCR Mix, 4x
MDX082
50 mL
10,000 Rxns
Liquid | Air-dried
Air-Dryable ™ RT-qPCR Mix | Applications:
PRODUCT
CAT NO. VOLUME REACTIONS
5 mL
1,000 Rxns
0% UTM+Sputum 5% UTM+Sputum 10% UTM+Sputum
Air-Dryable ™ 1-Step RT-qPCR Mix, 4x
MDX095
50 mL
10,000 Rxns
PRODUCT HIGHLIGHTS High-tolerance to PCR inhibitors in sputum samples
Amplification profile of a mouse RNA target spiked into samples containing 10%, 5% or 0% artificial sputum in Universal Transport Media (UTM). The data illustrates that the performance of Air-Dryable 1-Step RT-qPCR Mix (MDX095) exhibits high tolerance towards inhibitors present in artificial sputum and UTM.
HOW IT WORKS
*(Liquid mix can be dispensed into vials, plates or strips & subsequently air-dried under temperature-controlled conditions)
Air-Dryable ™ Direct DNA qPCR Blood |
Applications:
• Designed for the direct detection from whole blood, plasma and serum • Specimens require minimal sample processing, no extraction required PRODUCT HIGHLIGHTS
PRODUCT
CAT NO. VOLUME REACTIONS
5 mL
1,000 Rxns
Air-Dryable ™ Direct DNA qPCR Blood, 4x
MDX092
50 mL
10,000 Rxns
Serum
Plasma
High reaction efficiency on plasma, serum and whole blood samples containing anticoagulants Plasmid DNA containing Epstein-Barr virus was spiked into 10%serum or 10%K2-EDTA plasma and amplified in a triplex reaction using air-dried MDX092 format ( red ) and kits from Probe Force ( green ) and TaqPath ProAmp ( black ). The results illustrate higher end florescence and better sensitivity withMDX092 than with mixes from supplier R and T.
Air-dried | Probe Force | TaqPath ProAmp
Air-Dryable ™ Direct RNA/DNA qPCR Blood |
Applications:
PRODUCT HIGHLIGHTS
PRODUCT
CAT NO. VOLUME REACTIONS
High reaction efficiency with plasma, serum or whole blood samples containing anticoagulant K2-ETDA Dengue virus was amplified in a quadruplex reaction using air-dried MDX121 ( red ) and liquid mixes from Probe Force ( green ) and TaqPath ( black ) in the presence of 5% K2-EDTA blood or 5% K2-EDTA plasma. Air-Dryable ™ Direct RNA/DNA qPCR Blood has higher multiplexing capacity than mixes from supplier R and T.
5 mL
1,000 Rxns
Air-Dryable ™ Direct RNA/ DNA qPCR Blood
MDX121
50 mL
10,000 Rxns
Blood
Plasma
MDX121 (Air-dried) | Probe Force | TaqPath
www.meridianlifescience.com
Air-Dryable
Air-Dryable ™ Direct DNA qPCR Saliva |
Applications:
• Designed for the direct detection from sputum or saliva samples • Specimens require minimal sample processing, no extraction required PRODUCT HIGHLIGHTS
PRODUCT
CAT NO. VOLUME REACTIONS
5 mL
1,000 Rxns
Air-Dryable ™ Direct DNA qPCR Saliva
MDX130
50 mL
10,000 Rxns
Sensitive detection in multiplex assays using saliva samples (up to 60% human saliva) or Universal Transport Media (up to 35% UTM) Mycoplasma pneumoniae genomic DNA, adenovirus (ADV) and cytomegalovirus (CMV) were amplified in a triplex qPCR assay in the presence of 0% ( black ), 10% ( red ), 20% ( blue ) and 60% ( grey ) homogenized human saliva. The results illustrate that the performance of the air-dried, Air-Dryable Direct DNA qPCR Saliva is not affected by increasing concentrations of human saliva.
Mycoplasma pneumoniae
ADV
CMV
0% Saliva | 10% Saliva | 20% Saliva | 60% Saliva
Air-Dryable ™ Direct RNA/DNA qPCR Saliva |
Applications:
PRODUCT HIGHLIGHTS
PRODUCT
CAT NO. VOLUME REACTIONS
Sensitive detection inmultiplex assays using saliva samples (up to 60% human saliva) or Universal Transport Media (up to 35% UTM)
5 mL
1,000 Rxns
Air-Dryable ™ Direct RNA/DNA qPCR Saliva
MDX131
50 mL
10,000 Rxns
Three respiratory pathogens, Influenza A, Middle East Respiratory syndrome coronavirus (MERS-CoV) and Respiratory Syncytial Virus (RSV) were amplified in a triplex qPCR assay in the presence of 35% Universal Transport Media (UTM) with artificial sputum swab. The results illustrate that a higher performance was achieved with Air-Dryable ™ Direct RNA/DNA qPCR Saliva ( red ) compared to an inhibitor-tolerant RT-qPCR mix from TaqPathRT-qPCR Mix ( black ) and UltraPlex1-Step Tough Mix ( grey ).
Influenza A
MERS-CoV
RSV
Air-dried | TaqPathRT-qPCR Mix | UltraPlex1-Step Tough Mix
Air-Dryable ™ Direct DNA qPCR Stool |
Applications:
• Designed for the direct detection from stool • Specimens require minimal sample processing, no extraction required
PRODUCT
CAT NO. VOLUME REACTIONS
5 mL
1,000 Rxns
Air-Dryable ™ Direct DNA qPCR Stool
MDX140
50 mL
10,000 Rxns
PRODUCT HIGHLIGHTS
Sensitive detection in multiplex assays in the presence of stool inhibitors
E. coli O157 (Shiga-like/Verotoxin 2 gene), Salmonella typhimurium (16S-23S rRNA gene internal transcribed spacer) and Campylobacter jejuni (16S-23S rRNA gene internal transcribed spacer) were amplified in a triplex reaction using Air-Dryable ™ Direct DNA qPCR Stool ( red ) or mixes from Probe Force ( green ) or PerfeCTa qPCR ToughMix ( grey ) in the presence of 2 mg/mL bile salts. The results illustrate that Air-Dryable Direct DNA qPCR Stool is not affected by increased concentrations of bile, unlike Probe Force and PerfeCTa qPCR ToughMix.
Esherichia coli 0157
Salmonella typhimurium
Campylobacter jejuni
Air-dried | Probe Force | PerfeCTa qPCR ToughMixQ
Air-Dryable ™ Direct RNA/DNA qPCR Stool |
Applications:
PRODUCT HIGHLIGHTS
PRODUCT
CAT NO. VOLUME REACTIONS
Sensitive detection inmultiplex assays in the presence of stool inhibitors
5 mL
1,000 Rxns
Air-Dryable ™ Direct RNA/DNA qPCR Stool
MDX141
50 mL
10,000 Rxns
Two RNA targets (Rotavirus and Norovirus) and two DNA targets
Rotavirus
Adenovirus
(Adenovirus and Campylobacter jejuni ) were amplified in a quadruplex reaction using Air-Dryable ™ Direct RNA/DNA qPCR Stool ( red ) or mixes from TaqPath 1-Step RT-qPCR Master Mix ( black ) or UltraPlex 1-Step ToughMix ( grey ) in the presence of 1.5 mg/mL bile salts. The results illustrate that Air-Dryable Direct RNA/DNA qPCR Stool is less affected by increased concentrations of bile, unlike TaqPath 1-Step RT-qPCR Master Mix and UltraPlex 1-Step ToughMix.
Norovirus
Campylobacter jejuni
Air-dried | TaqPath 1-Step RT-qPCR Master Mix | UltraPlex 1-Step ToughMix
www.meridianlifescience.com
Air-Dryable
Air-Dryable ™ Direct DNA qPCR Urine |
Applications:
• Designed for the direct detection of bacteria and cell-free nucleic acids at very low titers from crude urine samples • Specimens require minimal sample processing, no extraction required
PRODUCT
CAT NO. VOLUME REACTIONS
5 mL
1,000 Rxns
Air-Dryable ™ Direct DNA qPCR Urine
MDX150
50 mL
10,000 Rxns
PRODUCT HIGHLIGHTS
Superior reaction efficiency with crude urine samples in a multiplex reaction
Neisseria gonorrhoeae
Chlamydia trachomatis
GBS
MDX150 (Air-dried) | PROBE FORCE | PerfeCTa qPCR ToughMix
Targets from Neisseria gonorrhoeae, Chlamydia trachomatis and group B streptococcus (GBS) were amplified in a triplex reaction in the presence of 25% human urine, using air-dried Air-Dryable ™ Direct DNA qPCR Urine ( red ) and PROBE FORCE ( green ) and PerfeCTa qPCR ToughMix ( grey ). The results illustrate higher multiplexing capacity and much better sensitivity using air-dried Air-Dryable Direct DNA qPCR Urine in the presence of 25% human urine.
Air-Dryable ™ Direct RNA/DNA qPCR Urine |
Applications:
PRODUCT HIGHLIGHTS
PRODUCT
CAT NO. VOLUME REACTIONS
Superior reaction efficiency with crude urine samples in a multiplex reaction
5 mL
1,000 Rxns
Air-Dryable ™ Direct RNA/DNA qPCR Urine
MDX151
50 mL
10,000 Rxns
Dengue
Zika
Chikungunya
MDX151 (Air-dried) | Reliance 1-Step Multiplex RT-qPCR Supermix | UltraPlex 1-Step ToughMix
Three viral RNA targets (Dengue, Zika and Chikungunya) were amplified in a triplex reaction using air-dried Air-Dryable ™ Direct RNA/ DNA qPCR Urine ( red ) and kits from Reliance 1-Step Multiplex RT-qPCR Supermix ( green ) and UltraPlex 1-Step ToughMix ( grey ) in the presence of 10% human urine. The results illustrate that dry Air-Dryable Direct RNA/DNA qPCR Urine has higher multiplexing capacity and reproducibility in the presence of 10% urine.
Air-Dryable ™ DNA LAMP Mix |
Applications:
• Fast reaction kinetics – results in half the time • Complete, pre-formulated mix for isothermal amplification of DNA and RNA targets – just add custom primers and probes • Ideal for POC diagnostic platforms or automated high-throughput instruments • Concentrated 4x formulation with flexibility in final assay format (liquid or air-dried) PRODUCT HIGHLIGHTS
PRODUCT
CAT NO. VOLUME REACTIONS
5 mL
800 Rxns
Air-Dryable ™ DNA LAMP Mix
MDX119
50 mL
8,000 Rxns
Results in half the time – Fastest reaction kinetics on the market
Air-Dryable ™ RNA/DNA LAMP mix was air-dried with Dengue type 2 primers at 50°C for 80 mins. The sensitivity of air-dried Air-Dryable ™ RNA/DNA LAMP mix ( red ) was compared to its liquid ( blue ) format and NEB WarmStart ® ( orange ) in a serial dilution of the target RNA (Dengue type 2 crude lysate). Reactions were incubated at 65°C for 60 min and TTR were measured at 1:10 of end fluorescence. Error bars represent standard deviation over 4 technical replicates. The results illustrate that the speed of Air-Dryable ™ RNA/DNA LAMP mix, is twice as fast as a leading supplier N mix in either the dry or liquid format.
40 30 20 10 0
3 ng
1.5 ng
0.3 ng
0.033 ng
MDX119 (Air-dried) | Liquid format | NEB WarmStart ®
Air-Dryable ™ RNA/DNA LAMP Mix |
Applications:
PRODUCT HIGHLIGHTS
PRODUCT
CAT NO. VOLUME REACTIONS
High reaction specificity with little-to-no non-specific amplification
5 mL
800 Rxns
Air-Dryable ™ RNA/DNA LAMP Mix
MDX118
50 mL
8,000 Rxns
Air-Dryable ™ RNA/DNA LAMP (MDX118) ( blue ) was compared to NEB WarmStart ® ( orange ) for detection of EBV DNA (1000 ipc) and MS2 RNA (1000 ipc). Reactions were incubated at 65°C for 60 min and TTR were measured at 1:10 of end fluorescence. Amplification plots of 3 technical replicates show Air-Dryable ™ RNA/DNA LAMP mix has earlier TTR and no non-specific amplification compared to supplier N which has detectable amplification for non-template controls (NTC).
EBV-DNA
MS2-RNA
MDX118 | NEB WarmStart ®
Lyo-Ready
• Ready-to-use, glycerol-free qPCR master mix formulated with a specialized blend of excipients • Ideal for multiplex assays on automated high-throughput instruments • Compatible with a range of lyophilization protocols to produce freeze-dried beads or cakes PRODUCT FEATURES:
Lyo-Ready™ qPCR Mix | Applications:
• Sensitive detection in multiplex assays with low-copy number targets • Virus mix (MDX062) is optimized for amplification of RNA or DNA viruses with a high secondary structure from either extracted or intact virus samples
PRODUCT
CAT NO. VOLUME REACTIONS
5 mL
500 Rxns
Lyo-Ready ™ qPCR Mix
MDX021
100 mL 10,000 Rxns
4 mL
500 Rxns
Lyo-Ready ™ qPCR Mix 2.6x
MDX023
100 mL 12,500 Rxns
PRODUCT HIGHLIGHTS Powerful multiplexing capacity
Three viral sequences, Cytomegalovirus (CMV) Adenovirus, Epstein-Barr Virus (EBV) and a DNA Extraction Control were amplified with equal efficiency from synthetic DNA templates with Lyo-Ready qPCR Mix in a quadruplex qPCR probe assay.
CMV
EBV
Adenovirus Amplification Plot
DNA Extraction Control
Amplification Plot
Amplification Plot
Amplification Plot
Cycle
Cycle
Cycle
Cycle
Lyo-Ready ™ 1-Step RT-qPCR Mix | Applications:
PRODUCT
CAT NO. VOLUME REACTIONS
PRODUCT
CAT NO. VOLUME REACTIONS
10 mL
1,000 Rxns
10 mL
1,000 Rxns
Lyo-Ready ™ 1-Step RT-qPCR Mix
Lyo-Ready ™ 1-Step RT-qPCR Virus Mix
MDX024
MDX062
100 mL 10,000 Rxns
100 mL 10,000 Rxns
PRODUCT HIGHLIGHTS High sensitivity amplification from both DNA and RNA templates Rotavirus A (dsRNA) and Adenovirus (dsDNA) were amplified in a single multiplexed RT-qPCR assay using inactivated crude viral lysates and Lyo-Ready 1-Step RT-qPCR Mix. The result illustrates that both viruses were amplified with high sensitivity, demonstrating the ability of Lyo-Ready 1-Step RT-qPCR Mix to detect low-copy number RNA and DNA targets simultaneously from a single sample.
Adenovirus Rotavirus A
Cycle
HOW IT WORKS
Key Steps in Lyophilization Process
Dried product in cake or bead form
Solution is frozen solid
Frozen solution undergoes primary (Sublimation) and
Plates, vials or strips are filled with solution
secondary (Desorption) drying processes under vacuum
Lyo-Ready ™ LAMP Mix | Applications:
• Ideal for POC diagnostic platforms or automated high-throughput instruments
PRODUCT
CAT NO. VOLUME REACTIONS
5 mL
800 Rxns
• Optimized for isothermal amplification such as Loop Mediated Isothermal Amplification (LAMP) • Concentrated 4x master mix • Contains all the required excipients for subsequent lyophilization PRODUCT HIGHLIGHTS Better Sensitivity with Lower Sample Input
Lyo-Ready™ LAMP Mix
MDX097
50 mL
8,000 Rxns
MDX097 | Isothermal Mastermix | NEB WarmStart ®
20
The average time to results (TTR) for Lyo-Ready ™ LAMP Mix ( blue ) and a mix from Isothermal Mastermix ( tan ) and NEB WarmStart ® ( orange ) were compared using a 10-fold serial dilution for the of BRCA1 gene (1,000, 100 and 10 copies). The relative concentrations of LAMP oligos were optimized to obtain earlier TTR compared to other experiments. The results demonstrate the increased sensitivity and greater reproducibility of the Lyo-Ready LAMP Mix, with earlier TTR values and a lower SD when compared to other suppliers. Reactions were incubated at 65°C for 60 min and TTR were measured at 1:10 of end fluorescence.
15
10
5
0
1,000 copies
100 copies
10 copies
www.meridianlifescience.com
Lyo-Ready
Lyo-Ready ™ RT-LAMP 1-Step Mix | Applications:
PRODUCT HIGHLIGHTS Quicker Time to Results and Better Sensitivity in the Presence of Inhibitors
PRODUCT
CAT NO. VOLUME REACTIONS
5 mL
800 Rxns
Lyo-Ready™ RT-LAMP 1-Step Mix
MDX108
50 mL
8,000 Rxns
A) UTM and sputum
B) Saliva
60
60
50
50
40
40
30
30
20
20
10
10
0
0
0% UTM + Sputum
10% UTM + Sputum
20% UTM + Sputum
0% Saliva
20% Saliva
25% Saliva
MDX108 | NEB WarmStart ®
A Dengue Type 2 RNA target was amplified from a 1:10 dilution of Dengue Type 2 crude extract in the presence of different concentrations of a (A) UTM and artificial sputum and (B) artificial saliva using Lyo-Ready ™ RT-LAMP 1-Step Mix ( blue ) and NEB WarmStart ® ( orange ). Reactions were performed at 65°C for 60 min in 4 replicates. Time-to-result is set as the time at which the fluorescence crossed a threshold of 10% of maximal fluorescence. The results illustrate the faster average time-to-result (TTR) with the Lyo-Ready ™ RT-LAMP 1-Step Mix, demonstrating faster detection without inhibitors and less inhibition at high concentrations of both UTM and sputum and saliva compared to NEB WarmStart ® mix.
DNA Polymerase
Aptamer Taq HS (Glycerol Free) |
Applications:
• Highly suited to multiplex, high-throughput viral detection assays requiring high specificity • No activation step - reduce an assay run time by up to 15 minutes • Convenient room temperature reaction set-up • High enzyme concentration (50 U/μL) compatible with lyophilization protocols
PRODUCT
CAT NO. VOLUME REACTIONS
Aptamer Taq HS (glycerol free) (50 U/μL)
20 µL
50 mL
MDX015
500 µL
500 mL
PRODUCT HIGHLIGHTS Fast Hot-Start
Comparison of qPCR performance of Aptamer Taq HS (glycerol free) vs an antibody hot-start Taq. PCR reactions were run with and without an initial 2 minute high temperature activation step for both polymerases and Ct values were compared. No difference was observed in the Ct values between the reactions for the Aptamer Taq HS (glycerol free), indicating that an activation step does not have an effect on polymerase performance. In contrast, the antibody hot-start Taq had a lower Ct value in the reaction with a 2 min high-temperature denaturation step indicating a improvement in performance over the reaction with no activation step. Overall, the data illustrates the immediate activation of the Aptamer Taq HS (glycerol free) allowing for faster hot-start and faster reaction protocols.
Condition
Glycerol-Free Taq HS 50 U/µL |
Applications:
• Lyophilization-compatible DNA polymerase ideal for automated high-throughput testing • Provided as separate ‘tubes’ of Taq, hot-start antibody, and enzyme dilution buffer • Can be used with Lyo-Ready qPCR Buffer (Cat #MDX022) to develop lyophilized qPCR mixes
PRODUCT
CAT NO. VOLUME REACTIONS
20 µL
1,000 Units
Glycerol-Free Taq HS 50 U/µL
MDX011
500 µL 25,000 Units
4 mL
1,000 Rxns
Lyo-Ready qPCR Buffer 2.5x
MDX022
100 mL 12,500 Rxns
PRODUCT HIGHLIGHTS Comparison of freshly prepared and
lyophilized glycerol-free Taq HS master mixes A 10-fold serial dilution of template DNA was used to set up two sets of qPCR assays. One set was lyophilized ( blue ) and the other left as a liquid mix ( black ). Results illustrate that lyophilization of Glycerol-Free Taq HS does not have an effect on the quality of the qPCR, as both the assays demonstrate identical efficiency and sensitivity.
Cycle
www.meridianlifescience.com
Reverse Transcriptase
Lyo-compatible MMLV-RT |
Applications:
• High-concentration MMLV-RT suitable for incorporation into lyophilized RT-PCR assays • Demonstrates high efficiency in RT-qPCR assays and high sensitivity detection of low copy number RNA targets
PRODUCT
CAT NO. VOLUME REACTIONS
8 µL
1,000 Rxns
Lyo-compatible MMLV-RT
MDX042
80 µL
10,000 Rxns
55C MMLV-RT |
Applications:
• Reverse transcriptase activity up to 60°C • Ideal for RNA with high secondary structure such as viral genomes • Sensitive detection of low copy number RNA targets • Formulation compatible with lyophilization and air-drying applications
PRODUCT
CAT NO. VOLUME REACTIONS
50 µL
10,000 Units
200 µL
40,000 Units
55C MMLV-RT
MDX117
1 mL
200,000 Units
10 mL
2,000,000 Units
To test a sample or to ask additional questions, E: info@meridianlifescience.com
Ordering information: USA 5171 Wilfong Road
Email: info@meridianlifescience.com Orders: orders@meridianlifescience.com www.MeridianLifeScience.com
Memphis, Tennessee 38134 Phone: +1 901-382-8716 Fax: +1 901-333-8223
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11/21
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