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.
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Latest Insights and Updates | Q3 2024
• Product Spotlight: - TetroScript Reverse Transcriptase • replace SuperScript with a cheaper and better alternative! - SensiFAST ™ Probe Direct SuperMix • newest mix to the portfolio, includes new FAQs! - SimpliFi HS Mix • highest fidelity on the market! • Webinars: - The evolution of STI at-home testing solutions: Exploring key advancements in molecular and immunoassay testing for STI at-home kits IN THIS EDITION ：
Editor’s Notes
Dear Colleagues, Welcome to Meridian Biosciences’ Distributor newsletter where we will provide the latest news, product education, webinars, and additional resources etc to keep you up to date. As a life science manufacturer, we develop, manufacture, and distribute a broad range of innovative molecular mixes, enzymes and buffers for researchers. We remain dedicated to developing the best solutions for molecular research, that give answers with speed, accuracy, and simplicity. In this edition, we present to you our new, TetroScript Reverse Transcriptase, SensiFAST™ Probe Direct SuperMix and SimpliFi HS Mix. We provide updates on our recently published webinar titled “ The Evolution of STI At-HomeTesting Solutions ”. Don’t miss the chance to meet with us at several upcoming conferences, including NexGen Dx, ASHG and MEDICA! Links to schedule a meeting are available at the end of this newsletter. We also invite you to join us for our presentations at NextGen Dx: “ From High-Throughput to Point-of- Care OncologyTesting: Cutting-Edge Genotyping Chemistries “ and “ Increasing Access to Point-of- Care Diagnostics through Innovative Advances in Molecular Reagents for Assay Development ”. We look forward to seeing you soon!
• Resources:
- Distributor portal - Ciloo
• Meet us at an upcoming conference: - ADLM: July 30th – August 1st - NextGen DX: August 19th -21st - ASHG: Sep 4th -6th
• Common Questions & Answers
The MarketingTeam
www.bioline.com
| 1 Quarterly newsletter
Product Spotlights
TetroScript Reverse Transcriptase The only Reverse Transcriptase with comparable performance to SuperScript
TetroScript Reverse Transcriptase is a high concentration, robust thermostable Moloney Murine Leukemia Virus- Reverse Transcriptase (MMLV-RT), designed to reduce RNase H activity and increase thermal stability. The enzyme offers higher cDNA yields and improved efficiency on GC-rich target RNAs and RNA with high secondary structure, such as viral RNA. Key Features: • High thermostability: reverse transcriptase activity up to 60°C, ideal for RNA with high secondary structure such as viral genomes • Higher enzyme efficiency and sensitivity: reliable quantification of low abundance targets and scarce samples • Low glycerol: formulation compatible with lyophilization and air-drying applications • Fast: delivers reproducible, multiplex accurate RT-qPCR assay results in as little as 30 minutes
TetroScript vs Superscript – thermostability comparison
Performance of TetroScript Reverse Transcriptase ( red ), standard MMLV-RT ( grey ) and SuperScript III (ThermoFisher) ( black ) after pre-incubation at 40°C to 70°C for 10 mins, in a multiplex one-step RT-qPCR assay. Δct values were calculated against the ct value produced by the same enzyme stored at-20 °C. The data illustrates the increased thermal stability of TetroScript Reverse Transcriptase when compared to other MMLV-RT enzymes and its ability to efficiently synthesize cDNA at temperatures up to 60°C.
Compatible with lyophilization and air-drying TetroScript Reverse Transcriptase proprietary formulation allows for incorporation into assays designed for subsequent lyophilization.
Retains the same activity after drying
Air-dried | Wet
TetroScript Reverse Transcriptase was used in an air-dryable RT-qPCR Mix and dried down in a fan assisted oven ( red ). After rehydration, the mix was tested against freshly prepared liquid mix ( blue ), in a triplex one-step RT-qPCR assay on respiratory RNA virus targets (Influenza A, MERS-CoV and RSV). The results demonstrate that TetroScript Reverse Transcriptase retains the same activity level after air-drying even in challenging conditions such as multiplex RT-qPCR reactions.
Applications: The high-quality first-strand cDNA generated makes TetroScript Reverse Transcriptase ideal for: • Construction of cDNA libraries • 2-Step qPCR assays • Viral RNA targets • Gene cloning
PRODUCT
CAT NO.
REACTIONS
BIO-65111 10,000 Units BIO-65112 4 x 10,000 Units
TetroScript Reverse Transcriptase (200 U/μL)
To review the product in detail or download our flyer, visit: https://www.bioline.com/thermostable-reverse-transcriptase.html
www.bioline.com
| 2 Quarterly newsletter
SensiFAST TM Probe Direct SuperMix Advanced mix for direct DNA amplification from crude samples without extraction SensiFAST™ Probe Direct SuperMix is a highly inhibitor-resistant qPCR master mix that provides quick and easy extraction and amplification of DNA from a variety of tissue types. The SuperMix maximizes sensitivity while simultaneously minimizing the effect of blood, tissue and plant PCR inhibitors, to deliver greater experiment success rates. Key Features: • Inhibitor tolerant: optimized proprietary buffer system designed for overcoming common PCR inhibitors in crude lysate or unprocessed blood, tissue and plant samples • Specific: antibody-mediated hot-start DNA polymerase minimizes non-specific amplification for improved assay sensitivity and reliability • Sensitive: reliable quantification of low abundance targets and scarce samples • Reproducible: consistent results between technical replicates for increased confidence in results • Fast: delivers reproducible, multiplex accurate assay results in as little as 30 minutes High qPCR efficiency across a range of inhibitors SensiFAST™ Probe Direct SuperMix is a combination of the latest advances in buffer chemistry and PCR enhancers and stabilizers, together with an antibody-mediated hot-start polymerase, dNTPs and MgCl 2 . It has been designed for highly reproducible, accurate assay results in the presence of inhibitors, making it ideal for direct amplification from the most challenging samples. SensiFAST™ Probe Direct SuperMix has been tested with a variety of crude samples known to inhibit PCR, from whole blood to proteinase lysed crude DNA extracts, making it ideal for everything from biofluids to animal and environmental samples.
Efficient amplification from different sample types
SensiFAST Probe Direct SuperMix | Roche
Applications: Eliminating costly, time-consuming, cumbersome purification procedures that can lead to sample loss, not only increase the speed and sensitivity of the assay, but also makes SensiFAST™ Probe Direct SuperMix especially valuable for high-throughput applications with limited starting material, such as gene expression experiments, making it ideal for A 10-fold serial dilution of genomic DNA was spiked into cerebrospinal fluid, human urine and cow whole milk was amplified, using SensiFAST™ Probe Direct SuperMix ( green ) and Kapa Probe Force (Roche) ( red ) and the manufacturers’ recommended protocol. The results illustrate Sensi- FAST™ Probe Direct SuperMix is more sensitive than Kapa Probe Force, as lower dilutions could be detected, with better efficiencies for all three inhibitor samples.
• Gene expression analysis • Viral and bacterial detection • GMO testing • SNP genotyping • Mutation detection • Environmental monitoring
PRODUCT
CAT NO.
REACTIONS
BIO-86105 500 Reactions BIO-86120 2,000 Reactions
SensiFAST™ Probe Direct SuperMix
To review the product in detail or download our flyer, visit: https://www.bioline.com/sensifast-probe-direct-supermix.html
www.bioline.com
| 3 Quarterly newsletter
What sample types can be used with the Inhibitor- Tolerant qPCR Mixes? We have tested whole human and bovine blood, saliva, sputum, urine, cerebrospinal fluid (CSF), cow milk, stool sample, and plant lysate; other types of samples can be used but we would recommend testing different concentrations to determine the concentrations that can be tolerated. Do I need to add additional magnesium to the reaction? The SensiFAST™ Probe Direct SuperMix has been optimized for magnesium, dNTPs, stabilizers, and enhancers, resulting in highly reproducible, accurate RNA and DNA target amplification under fast thermal cycling conditions, delivering excellent results in qPCR multiplex assays, even in the presence of difficult PCR inhibitors. So, we do not recommend adding any additional material, only the primers, probe(s), and sample. SensiFAST TM Probe Direct SuperMix
PRODUCT FAQs
What are the recommended reaction conditions for qPCR multiplex assays? When multiplexing, standard conditions can be used, however, if necessary, the reverse transcription reaction time can be extended up to 20 minutes and/or the temperature can be increased up to 55° C and the annealing/extension time can be extended up to 60 seconds and/or the temperature can be increased up to 65° C, but this will need to be tested during optimization. Where do I begin with optimization of crude samples or crude extracts? We would recommend using a range of crude sample concentrations or extract volumes to ascertain at what point unacceptable PCR inhibition starts to take place. This will vary as some sample types, such as blood and stool specimens typically contain more inhibitors for qPCR/RT- qPCR reactions.
SimpliFi HS Mix Optimised for NGS library amplification with low GC bias
SimpliFi HS Mix is a high-fidelity polymerase mix using aptamer-based hot-start technology, highly suited to amplification of DNA from crude samples, for fast, inexpensive target enrichment, NGS library amplification and cloning applications.
Key Features: • Robust: optimized enzyme/buffer mix promotes reliable amplification of a broad range of targets, including complex DNA extracted from human, animal and plant samples • Specific: the aptamer-based hot-start remains completely inactive during PCR set-up and after amplification, to prevent non-specific products • Higher yield and sensitivity: high yields with minimal optimization, regardless of a template’s GC content, reducing time to results and eliminating the cost of unnecessary repeats • Enhanced accuracy: higher than 130x Taq fidelity, reducing errors for next generation sequencing (NGS) library amplification. High fidelity SimpliFi HS Mix is a combination of the latest advances in buffer chemistry and PCR enhancers and stabilizers, together with an aptamer-mediated hot-start polymerase, dNTPs and MgCl 2 . It has been designed for highly reproducible, accurate assay results in the presence of inhibitors. The advanced buffer chemistry and enhancers has been developed for fast PCR and is designed for superior sensitivity and specificity, making SimpliFi HS Mix perfect for NGS library amplification.
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| 4 Quarterly newsletter
SimpliFi HS Mix is a high-fidelity, high-efficiency hot-start PCR master mix for NGS workflows requiring DNA library amplification prior to sequencing 1 . The fidelity of the SimpliFi HS Mix was determined using single-molecule sequencing and compared to commercially available high-fidelity master mixes. The results are demonstrating the exceptional fidelity of SimpliFi HS Mix, offering higher level of confidence for preserving DNA sequence accuracy during the preparation of DNA for next-generation sequencing.
Fidelity comparison across commercially available high-fidelity polymerases
Purified plasmid DNA was extended using SimpliFi, the complementary strands were synthesized using a standard high-fidelity polymerase, in both cases using primers containing a partial Illumina adapter, a random product barcode and a condition barcode. Primers complementary to the partial Illumina adapter are used to PCR amplify the complementary strands, forming the sequencing library. After next-generation sequencing, reads are grouped according to condition barcode and product barcode. Sequences are aligned to the correct sequence and errors are called. Errors are only kept if they are present in all copies, otherwise they are discarded as sequencing error. Exactly the same method was used to determine the error rate for Q5 (NEB), Kapa HiFi HotStart ReadyMix (Roche) and Phusion HS Flex (NEB), and the fidelity values were normalized to Taq polymerase fidelity.
Optimized for low GC bias PCR amplification can be susceptible to bias resulting from genomes that contain unusually high or low GC content, SimpliFi HS Mix has been developed to address this through careful selection of high-quality polymerase and optimization of the buffer, making it ideal for NGS library preparation.
GC bias with both high and low GC content
Libraries were prepared from 100 ng (amplified libraries) or 1 μg (PCR-free library) of genomic DNA from S. aureus (32% GC) and R. spheroides (69% GC), mixed equimolarly. End-repair and ligation steps were carried out using JetSeq™ ER and Ligation Kit (BIO-68026) and for all the samples. Library amplification consisted of 10 PCR cycles using SimpliFi HS Mix and Kapa HiFi HotStart ReadyMix (Roche), following the manufacturer’s recommendations. GC bias plots were generated, with %GC content of 100 bp windows on the X axis. Normalized coverage is indicated for the High- SimpliFi HS Mix ( blue line ) and Kapa HiFi ( green line ) and can be compared to a PCR-free reference library ( black line ), which has the horizontal bias distribution closer to 1 (representing unbiased coverage). The results illustrate SimpliFi HS Mix has a lower bias across the entire range of GC content, compared to the PCR- amplified library using Kapa HiFi HotStart ReadyMix (Roche) especially for GC percentages higher than 65%.
Supplier K | SimpliFi | PCR-free Reference
Applications: • Gene expression • Viral and bacterial detection • Robust PCR • High-specificity PCR
PRODUCT
CAT NO.
REACTIONS
BIO-25060 100 Reactions BIO-25061 500 Reactions
SimpliFi HS Mix (2x)
To review the product in detail or download our flyer, visit: https://www.bioline.com/simplifi-hs-mix-2x.html
• High-fidelity PCR • GC/AT-rich PCR
References 1. Lee D.F., et al. (2016) Mapping DNA polymerase errors by single-molecule sequencing. Nucleic Acids Res. 44(13): e118; PMID: 27185891
www.bioline.com
| 5 Quarterly newsletter
WEBINAR
The evolution of STI at-home testing solutions: Exploring key advancements in molecular and immunoassay testing for STI at-home kits
Watch this webinar recording to learn about the changing landscape of STI testing, highlighting the new developments in at-home testing and the molecular and immunoassay reagent solutions for developing new STI assays. With the global increase of sexually transmitted infections (STIs), the need for accessible and accurate STI diagnostic solutions has never been more critical. Meridian offers complete solutions for immunoassay and molecular diagnostics across a wide variety of STD targets such as HIV-1/HIV-2, Hepatitis B, Syphilis, HPV and Herpes. From high-performing antigens and antibodies to inhibitor-tolerant qPCR and LAMP master mixes, Meridian supports diagnostic assay developers with valuable raw materials optimized to elevate assay performance. Visit our STD resource page to learn more.
RESOURCES
Distributor Portal
Ciloo We are updating the resource centre to equip our distributors with the tools you need to learn about our products, including product flyers, training slides and training recordings. Please log in to the distributor portal to access the latest product flyers and training resources. Any questions, please contact alpha.chen@meridianlifescience.com. Please log in to the Ciloo platform to access the latest marketing materials. Select the flyers you need (you can insert your logo and contact information at the footer) and print them directly from the platform to ensure you have the most up-to-date information and promotions. If you encounter any issues, please contact deirdre.leavy@meridianbioscience.com.
www.bioline.com
| 6 Quarterly newsletter
Meet us at upcoming conferences!
ADLM July 30th – August 1st, Chicago IL, Booth# 351 Book a meeting here
NextGen DX August 19th-21st , Washington DC, Booth # 18 Book a meeting here
Talk #1: August 19th, 11:00am From High-Throughput to Point-of-Care OncologyTesting: Cutting-Edge Genotyping Chemistries Presented by Florent Chang-Pi-Hin, PhD (Meridan Bioscience VP of R&D, Life Science Division) Talk #2: August 19th, 4:45pm Enabling Point-of-Care Diagnostics ProgramTrack Presented by Lina Gasiunaite (Meridian Bioscience R&D Team Leader, Life Science Division)
More to come ：
ASHG (Association for Human Genetics) Sep 4th-6th, Denver, Booth #385
· How to submit my suggestion for the next quarterly newsletter? Please email alpha.chen@meridianlifescience.com. Questions & Answers
Ordering
EMEA Region: Reni Lam reni.lam@meridanlifescience.com APAC & LATAM Region: zoe.kim@meridianlifescience.com
· Who do I contact for a missing order? Please contract info@meridianlifescience.com.
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