RT-PCR RT-qPCR

Why Choose Challenger Hotstart RT?Ultra-Fast cDNA synthesis – Achievable in just 2 minutes.Extended stability – In pre-assembled all-in-one reaction mixtures, Challenger Hotstart RT maintains full performance even after 7 days at 37 °C.Room-temperature setup – Pipette without cooling, saving time and reducing errors.Warm-start reverse transcriptase – Activated only at high temperatures, preventing nonspecific cDNA synthesis before the reaction begins.Exceptional thermostability – Over 99 % of enzyme activity remains sealed at 42 °C for controlled and reliable activation.Versatile applications – Ideal for rapid cDNA synthesis, RT-PCR, and RT-qPCR with challenging RNA templatesOptimized for High-Performance Reverse Transcription Workflows Challenger Hotstart RT is designed for demanding applications. Its Warm-Start technology keeps the enzyme inactive during setup, enabling precise control over the start of cDNA synthesis. The improved thermostability ensures robust performance: Challenger Hotstart RT remains inactive at 42 °C and is only activated at higher temperatures. Therefore, a standard reverse transcription reaction with oligo(dT) primers or random hexamer primers at 42 °C is not possible with this enzyme.Fast and Reliable cDNA SynthesisChallenger Hotstart RT allows efficient reverse transcription even with templates that have complex secondary structures or high GC content. Its enhanced thermostability ensures rapid and accurate cDNA generation without compromising specificity or yield.Long-Term Stability and Easy HandlingEngineered for routine and high-throughput lab work, Challenger Hotstart RT supports room-temperature setup, simplifying workflows and reducing errors. Its robust formulation guarantees reliable storage and shipping stability.Typical ApplicationscDNA synthesis from total RNA or mRNA, including difficult templates.One-step or two-step RT-qPCR with high sensitivity, even at low RNA input.Research and diagnostic workflows requiring reproducible results.High-throughput applications, including multiplexed panels or screening assays.Key Features SummaryUltra-Fast cDNA synthesis – as quick as 2 minutes.Extended stability – retains full performance after 7 days at 37 °C in premixed reaction setups.Warm-start reverse transcriptase – delayed activation improves specificity.High thermostability – supports challenging RNA templates. Room-temperature setup – simplifies workflow, reduces errors. Robust performance – suitable for routine and high-throughput applications.

Why Choose Challenger Hotstart RT-qPCR Mix?Room-temperature setup – Easy pipetting without cooling, saving time and reducing errors.warm-start reverse transcriptase – Prevents nonspecific amplification and allows primer/probe premixing.Superior contamination control – UDG/dUTP system eliminates carryover contamination for reliable qPCR results.Fast RT-qPCR protocols – Complete assays in just 30 minutes with a 5-minute reverse transcription step.High sensitivity RT-qPCR – Detect as few as 1–10 RNA copies.Flexible applications – Single and multiplex RT-qPCR for viral detection, gene expression, and diagnostics.Optimized for High-Performance qPCR WorkflowsThe 4× concentrated formulation allows high-throughput and multiplexing while keeping reaction efficiency high. The combination of warm-start RTase and antibody-modified Taq DNA polymerase and a RNAse inhibitor ensures precise control, so your reactions only start when you are ready.Contamination-Free One-Step RT-qPCRBuilt-in dUTP/UDG safeguards protect your assays from false positives due to carryover contamination. This, in combination with an RNase inhibitor is essential for labs running high-sensitivity viral RNA detection and diagnostic assay developments.Fast RT-qPCR Protocols Without Sacrificing AccuracyWith Challenger, you can speed up detection while keeping sensitivity intact:Standard program: 55 °C for 15 min RT, ~1 hr totalFast program: 55 °C for 5 min RT, ~30 min total Benchmark studies with HBV/HCV RNA samples confirmed superior stability and consistent Ct values compared to competitor brands.Long-Term Stability You Can TrustUnlike conventional RT-qPCR mixes, the Challenger Hotstart RT-qPCR Master Mix is engineered for long shelf life. Even under accelerated testing at 37 °C for 10 days, the mix maintained stable multiplex detection - ensuring reliable performance during storage and shipping.Example ApplicationsViral RNA detection: HBV, HCV, Influenza, RSV, SARS-CoV-2, hMPV, PIV1, and moreGene expression analysis: accurate detection of high- and low-copy RNA targetsClinical research & diagnostics: robust, reproducible dataHigh-throughput testing: multiplex RT-qPCR panels for respiratory, gastrointestinal, and hepatitis virusesMany more…Key Features SummaryOne-step RT-qPCR Master Mix with warm-start RTaseHigh sensitivity qPCR mix – detects single RNA copiesFast RT-qPCR protocol – 5 min RT, 30 min totalLong stability RT enzyme – stable at 37 °C for 10 daysUDG/dUTP contamination control – eliminates carryover risk4× concentrated format – supports multiplexing High sensitivity and stable detectionStable detection of single-digit copy templates using the Challenger Hotstart RT-qPCR 4X Mastermix:

Our One-Step RT-qPCR 2X master mix was developed for quantitative real-time analyzes of RNA templates using SybrGreen®. The ready-to-use mixture is based on a genetically modified reverse transcriptase with improved thermal stability, which offers increased specificity, a high cDNA yield and improved efficiency for highly structured and long cDNA fragments.The 2X Mastermix contains all the reagents required for the RT-qPCR (except for the template and primer) to ensure quick and easy handling at a minimum of pipetting steps. The high-quality enzymes and the optimized reaction buffer with high-purity dNTPs guarantee superior real-time PCR results.This Master Mix does not contain ROX - please make sure to choose the right setting on your qPCR cycler. A list of common thermal cyclers and their ROX compatibility can be found under the "Tips" tab.RT-qPCR is used to amplify double-stranded DNA from single-stranded RNA templates to enable rapid real-time quantification of RNA targets. In the reverse transcription step, the reverse transcriptase synthesizes single-stranded DNA molecules (cDNA). The cDNA molecules are amplified by the HotStart DNA polymerase. The HotStart polymerase activity is blocked at ambient temperature and will be activated at the beginning of the initial denaturation. This thermal activation prevents the extension of non-specifically annealed primers and primer-dimer formations at low temperatures during the PCR setup.One-Step RT-qPCR offers enormous convenience when analyzing targets from multiple RNA samples and minimizes the risk of contamination. The Challenger Hotstart RT-qPCR 4x Mastermix (M3060 >) is particularly suitable for RNA that exhibits a high degree of secondary structures or for complementary primers! With our high-quality dNTPs, available as a set (M3015) or as a mix (M3016), as well as our DNA markers and affordable standard agarose, we offer you additional products for PCR.Time saving: up to 50%! No need for Superscript!Money saving: high!

Our One-Step RT-qPCR 2X master mix was developed for quantitative real-time analyzes of RNA templates using probe-based assays such as Taqman®, Beacons, MGB or Mediator Probes and can be directly used with RNA as a starting material. The ready-to-use mixture is based on a genetically modified reverse transcriptase with improved thermal stability, which offers increased specificity, a high cDNA yield and improved efficiency for highly structured and long cDNA fragments. The 2X Mastermix contains all the reagents required for the RT-qPCR (except for the template, primer and probes) to ensure quick and easy handling at a minimum of pipetting steps. The high-quality enzymes and the optimized reaction buffer with high-purity dNTPs guarantee superior real-time PCR results. The sophisticated buffer system provides fast kinetics, a RNA sensitivity of <10 fg and target amplification even for difficult templates and multiplexing applications for more than 6 targets.This Master Mix does not contain ROX - please make sure to choose the right setting on your qPCR cycler. A list of common thermal cyclers and their ROX compatibility can be found under the "Tips" tab.RT-qPCR is used to amplify double-stranded DNA from single-stranded RNA templates to enable rapid real-time quantification of RNA targets. In the reverse transcription step, the reverse transcriptase synthesizes single-stranded DNA molecules (cDNA). The cDNA molecules are amplified by the HotStart DNA polymerase. The HotStart polymerase activity is blocked at ambient temperature and will be activated at the beginning of the initial denaturation. This thermal activation prevents the extension of non-specifically annealed primers and primer-dimer formations at low temperatures during the PCR setup. One-Step RT-qPCR offers enormous convenience when analyzing targets from multiple RNA samples and minimizes the risk of contamination. The Challenger Hotstart RT-qPCR 4x Mastermix (M3060 >) is particularly suitable for RNA that exhibits a high degree of secondary structures or for complementary primers!With our high-quality dNTPs, available as a set (M3015) or as a mix (M3016), as well as our DNA markers and affordable standard agarose, we offer you additional products for PCR.Time saving: up to 50%! No need for Superscript!Money saving: high!

Ready to use One-Step RT-PCR  MasterMix - Standard mix.  Our RT-PCR One-Step Kit is designed for performing highly sensitive and specific RT-PCR. The kit is based on a genetically engineered reverse transcriptase with enhanced thermal stability resulting in an increased specificity, high cDNA yield and an improved efficiency for highly structured and long cDNA fragments. The kits contains all reagents required for RT-PCR (except template and primer) in one kit combining simple handling with high flexibility. The premium quality polymerases, ultrapure dNTPs and the optimized complete reaction buffers ensure superior amplification results. RT-PCR is used to amplify double-stranded DNA from single-stranded RNA templates. In the RT step the reverse transcriptase synthesizes single-stranded DNA molecules (cDNA) complementary to the RNA template. In the first cycle of the PCR step synthesis, Taq DNA polymerase synthesizes DNA molecules complementary to the cDNA, thus generating a double-stranded DNA template. During subsequent rounds of cycling the DNA polymerase exponentially amplifies the double-stranded DNA template.In one-step RT-PCR all components of reverse transcription (RT) and PCR are mixed in one tube prior to starting the reaction and thus carried out sequentially without opening the tube. This offers tremendous convenience when applied to analysis of single targets from multiple samples of RNA and minimizes the risk of contaminations.Genaxxon offers also a “Zero-Step” HotScriptase RT (M3056) a RNA/DNA Polymerase that enables direct PCR from RNA without an isothermal reverse transcription step. Just starting the PCR protocol with specific primers. Especially suited for RNA targets that exhibit a high degree of secondary structure, for self- or cross-complementary primers. The Challenger Hotstart RT-qPCR 4x Mastermix (M3060 >) is particularly suitable for RNA that exhibits a high degree of secondary structures or for complementary primers!

The Genaxxon M-MuLV Reverse Transcriptase, encoded by Moloney Murine Leukemia Virus (M-MuLV RT) and expressed in E.coli is an RNA-dependent DNA polymerase lacking the RNase H activity. The enzyme synthesizes the cDNA first strand from a single-stranded RNA template to which a primer has been hybridized. M-MuLV RT will also extend primers hybridized to single-stranded DNA! The enzyme is recommended for synthesis of cDNA from 100 bp up to 10 kb length. Second strand cDNA synthesis can be achieved from some mRNA templates without an additional DNA polymerase. Genaxxon offers the HotScriptase master mixes (M3062 >; M3064 >) with which it is possible to synthesize double stranded DNA completely without an isothermal transcription step. Time saving: up to 50%! No need for Superscript!Money saving: high! With our high quality dNTPs as Set (M3015.4100 and M3015.0250) or Mix (M3016.1010) or our DNA Ladders and our favourable standard agarose (M3044) we can offer additional high quality products for your PCR.

Genaxxon bioscience Scriptase RT is a proprietary reverse transcriptase with reduced RNase-H activity and increased thermostability that guarantees high specificity, longer cDNAs and a higher yield even with GC-rich target RNAs. With the included Oligo(dT) and Random Hexamer Primer you can easily optimise the protocol according to your needs.We recommend Scriptase RT for the synthesis of cDNA from 100 bp up to 10 kb in length.Properties Ready-to-use kit incl. primer and DNase Suitable for templates up to 17 kb High yield due to reduced RNase-H activity Complete reverse transcription in less than 15 minutes For specific primers, random hexamers or oligo(dT) primersreverse Transcription up to 65°C possible for complex transcripts Advantages High sensitivity cDNA in <15 minutes Primers included gDNA removal Temperature stability Applications  cDNA synthesis Fast and easy cDNA synthesis with just one kit. The Scriptase RT kit offers high sensitivity and specificity. It is also very flexible and delivers high yields with both oligo(dT) and random hexamers or gene-specific primers. Gene expression analysis Gene expression levels in tissues or different cell types can be determined by the detection of specific mRNA. For this purpose, the total RNA is first transcribed into cDNA. In the subsequent qPCR with gene-specific primers, the respective expression level is determined. cDNA sequencing (diagnostics) In the diagnosis of RNA viruses, reverse transcription is performed upstream of sequencing in order to create a cDNA library. In addition to pathogen detection itself, it can also be used to create mutation profiles of pathogens. RNA-seq As part of an RNA-seq workflow, mRNA enrichment can be performed using reverse transcription in combination with oligo(dT) primers. This allows both mRNA selection and RT to be performed in one step, which is particularly useful for small amounts of RNA. It should be noted that a 3′ bias may occur during subsequent sequencing. This would result in an increased number of reads for the 3′ end of the RNAs.

The Genaxxon RNase Inhibitor is a recombinant (E.coli) murine protein which inhibits ribonucleases (RNases) A, B and C. It does not inhibit RNase 1, RNase H, RNase T1, S1 Nuclease. Native RNase-Inhibitor from murine placenta exerts its inhibitory effect by binding non-covalently to RNases in a 1:1 ratio with high affinity. There is no inhibition of polymerase activity when this RNase Inhibitor is used with AMV Reverse Transcriptase, M-MuLV Reverse Transcriptase, Taq DNA Polymerase or Phage RNA Polymerase (SP6, T7, or T3).The enzyme is active over a broad pH range between 5 and 9, with a maximum activity at pH7 – pH8. The inhibition of RNases is a reversible process. Reagents as urea or thiol reagents will lead to the dissociation of the Inhibitor-RNase-complex, resulting in re-natured and functional RNase and irreversibly deactivated RNase inhibitor.Applications: RT-PCRcDNA synthesisIn vitro transcription/translationEnzymatic RNA labelling reactionsIn vitro virus detection. Improvement of RNA translation in homologous systems.Preparations of RNase-free antibodies. Any application where eukaryotic RNase contamination is a potential problem.  Advantages:  The recombinant murine RNase inhibitor does not contain the cysteine pair that was identified to cause the high sensitivity against oxidation resulting in the inactivation of the human RNase inhibitor (1). As a result, the murine RNase inhibitor has significantly improved oxidation resistance compared to the human and porcine RNase inhibitors. In addition, murine RNase inhibitor is much more stable at low low DTT concentrations (less than 1 mM), what makes it ideal for reactions where high DTT concentrations interfere with the reaction (e.g. real-time RT-PCR). Product does not contain glycerol => lyo-ready product (ready for preparations of freeze-dried products).Recommendations: Since the RNase inhibitor protein is sensitive to denaturing agents and conditions, whereas RNases are not, care must be taken to avoid high concentrations of urea or other denaturing agents and temperatures above 50-55°C. The recommended concentration of the RNase inhibitor is 1 unit/µL. During set-up of a reaction, RNase inhibitor should be added before other components are added (enzymes, plasmids from mini preps, etc.).References1. Kim, B.M. et al. Protein Science. 8, 430-434.

Random Hexamers are short oligodeoxyribonucleotides of random sequence [d(N)6]. Primers are quality controlled (MS), purified (reversed phase HPLC), lyophilised and aliquoted and came with a guaranteed amount of at least 5OD (about 76.1nmol (136.4µg)). Hexanucleotide primers are a mixture of random 5'-hydroxyl hexanucleotides or hexamers, and can be used to quickly and efficiently prepare radioactive or non-radioactive probes using a DNA polymerase and a suitable DNA template or for cDNA synthesis from mRNA. The heterogeneous nature of the random primers ensures that all possible sequences will be represented in the probe mixture. As a result, these primers are well suited for generating as long as possible (complete) transcripts of the RNA, transcribing the 5 'ends, in particular for long RNA templates. As an alternative you can also use Oligo(dT)-Primer > for reverse transcription. Oligo(dT) primer bind to the polyA tail of RNA and therefore transcription will start always at the 3'-end of the RNA. The guaranteed delivery quantity of 5OD, corresponding to approx. 76.1nmol, or 136.4μg, can be dissolved in 760μL DEPC water to give a concentration of 100pmol/μL (100μM), or approx. 180μg/μL. If using between 50ng and 250ng of the hexamers per 20μL reverse transcription reaction, 760μL (100μM) will be sufficient for 460 to 2280 reverse transcriptase reactions.

Oligo (dT)20 are single stranded oligodeoxyribonucleotides containing only deoxythymine (dT) to be able to prime with the poly(A) tail of mRNA molecules. Primers are quality controlled (MS), purified (reversed phase HPLC), aliquoted and lyophilized and came with a guaranteed amount of at least 5OD (about 27.2nmol (163.7µg)). Oligo (dT)20 Primer is designed to initiate the synthesis of cDNA from total RNA in a reverse transcription reaction, where Reverse Transcriptase, e.g. MMuLV (M3042) > is starting the reaction from the poly-A-end of mRNA. It can also be used for generation of labeled cDNA to screen microarrays. A mixture (1:1 ratio) of random hexamer primers (M3038 >) and Oligo(dT)20 primer may improve the sensitivity of cDNA synthesis as it will especially improve the efficiency of transcripts longer than 600 bp. Guaranteed delivery of at least 5OD (equals about 27.2nmol (163.7µg)). Primers are quality controlled (MS), purified (reversed phase HPLC), lyophilised and aliquoted. Dissolve content (5 OD) in 271.5μL of molecular biology grade water for an end concentration of 100 pmol/μL (100μM). 760μL (100μM) are sufficient for 760 reverse transcriptions of 20μL each. Oligo (dT)20 Primer is suitable for use as a primer for first strand cDNA synthesis with a reverse transcriptase, such as MMuLV or AMV. The primer hybridizes to the poly-adenylated tail found on the 3´ end of most eukaryotic mRNAs. Oligo (dT)20 ensures that the 3´ end of mRNAs are represented. Oligo(dT)20 Primer may be better suited for the new generation of Reverse Transcriptases like SuperScript™ that work at higher temperatures as the longer Oligo(dT)20 Primer enable annealing in reverse transcription reactions at higher temperatures. Applications• cDNA synthesis from total RNA in a reverse transcription reaction.• optimal choice for construction of cDNA libraries from eukaryotic mRNAs.• Full length cDNA cloning• 3’ rapid amplification of cDNA ends (3’ RACE)• Generation of labelled cDNA for screening of microarrays.Oligo (dT)20 Primer can not be used together with degraded RNA, prokaryotic RNA or miRNA (lack of poly(A) tail). Primer Sequence: 5´ – d (TTT TTT TTT TTT TTT TTT TT) –3´

The Genaxx1Step RT-qPCR SARS-CoV-2 CDC Probe Assay Kit is a real-time RT-PCR-based detection system for the targets N1 and N2 of the nucleocapsid from the SARS-CoV-2-Corona virus (2019-nCoV), as well as for the control target RNASeP. The kit has been optimized for qualitative in vitro detection according to the US CDC (Centers for Disease Control and Prevention).The Genaxx1Step RT-qPCR SARS-CoV-2 CDC Probe Assay Kit consists of the innovative HotScriptase Covid-19 master mix with its very thermostable polymerase. Primers and probes according to CDC-authorized sequences for the N1, N2 assays and for RNASeP can be purchased, for example, from IDT (Integrated DNA Technologies) or Eurofins. The test is not affected by following main variants (will detect these variants): BA 2.75, XBB 1.5. Genaxx1Step Covid-19 master mix enables the amplification of RNA or DNA target sequences with quick and easy PCR protocols, even without an isothermal reverse transcription step. The light blue color improves the visibility of the mixture during the pipetting steps, especially if white or transparent PCR plates or PCR tubes are used. faster: One-Step - NO time consuming isothermal intermediate step necessary. easier: reverse transcriptase and DNA polymerase in one single enzyme; reverse transcription and DNA amplification in parallel during the PCR elongation step. You need only a PCR protocol. saver: much reduced contamination risk, due to reduced pipetting steps. extreme thermostablility: secondary structures will be a minor problem due to higher temperatures of PCR compared to reverse transcription reactions. cheaper: parallel reverse transcription and amplification by HotScriptase RT saves time and costs. The Genaxxon HotScriptase DNA polymerase for reverse transcription enables a "One-Step” RT-PCR directly from cell suspensions or mammals cells without an isothermal reverse transcription intermediate step. Just give cells or cell suspension (100 up to <10,000 cells) directly to the PCR reaction mixture, mix, place in a thermal cycler and start PCR program. HotScriptase RT polymerase is a biotechnical engineered, extremely thermostable reverse transcriptase and combined DNA polymerase, obtained through directed, artificial evolution. The HotScriptase 2-time master mix is a ready-to-use mixture of the propietory HotScriptase, buffer, dNTPs and special components. The recommended amplicon size should be between 60-400 bp! This master mix contains no ROX nor green dye! > find out about our realtime master mixes