Master Mixes for Molecular Ambient-Temperature Stable Assays

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

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

Connect with us:

ISO 13485 Certified

02/22

Page 1 Page 2 Page 3 Page 4 Page 5 Page 6 Page 7 Page 8 Page 9 Page 10 Page 11 Page 12

meridianlifescience.com

Powered by