RT-PCR RT-qPCR - Products for reverse Transcription - RT-PCR - Covid-19 or SARS-CoV-2

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.

HotScriptase Probe Mastermix for RT-PCR without an isothermal intermediate step, thereby saving time and money: HotScriptase Probe Mastermix was successfully used by several customers for the detection of RNA viruses like Zika or SARS-CoV-2 (COVID-19 detection). HotScriptase is a extremely thermostable reverse transcriptase and DNA polymerase in one enzyme. HotScriptase Probe Mastermix 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 mastermixes

HotScriptase RT Master Mix for One-Step RT-PCR without isothermal reverse transcription step. Using HotScriptase RT is very time and cost efficienct as HotScriptase RT is a reverse transcriptase and a DNA Polymerase combined in one single highly thermostable enzyme. 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 a fluorescence dye but no ROX! > find out about our realtime master mixes

The One-Step RT-qPCR 2X ProbeMastermix without ROX® was developed for quantitative real-time analyzes of RNA templates with double-labelled fluorescence probes. The ready-to-use mixture is based on an optimized 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. 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. Genaxxon offers also 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!

Our One-Step RT-qPCR 2X master mix was developed for quantitative real-time analyzes of RNA templates using EvaGreen®. 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. As a further improvement, EveGreen® was used as the fluorescent dye, which inhibits PCR less than SYBR® Green. 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. 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. Genaxxon offers also 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!

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

Improved version of our kit M3068. Now coming as a 2-time ready-to-use RT-PCR master 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. Genaxxon offers also 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!

The Genaxx1Step one-step RT-qPCR SARS-CoV-2 Garble Kit contains our innovative HotScriptase RT which is an extremely thermo- and inhibitor stable DNA polymerase with reverse transcriptase activity which does not need purified RNA for transcription and amplification. An aliquot of a gargling test solution is enough for direct RT-PCR. Also unlike some of our competitor´s kits, Genaxx1step RT-PCR SARS-CoV-2 Gargle Kit can be used on commonly used real time PCR machines such as Lightcycler 480 or BioRad CFX.The HotScriptase polymerase facilitates an “OneStep” RT-PCRs directly from RNA templates, as reverse transcription takes place simultaneously with DNA amplification during the cycled PCR elongation step. Besides the very convenient protocol this allows reverse transcription reactions at higher temperatures, thus minimizing the problems encountered with strong secondary structures in RNA, or strong tertiary structures as in double strand RNA viruses that melt at elevated temperatures. High temperatures helps implementing a heat denaturation step sufficient to destroy the viral envelope and thus releasing the viral RNA. Furthermore, high temperatures help to destroy secondary structure structures in the viral RNA which are often found in single-stranded RNA viruses, e.g. pseudoknots, loops and hairpins, which represent a difficult target for other transcriptases than HotScriptase and can cause the enzyme to detach from the RNA strand. Efficient transcription is thus often not possible but in the case of our Covid-19 direct kit, a temperature of 75°C is applied during the transcription process, which eliminates such possible blocks. Now even better, easier and faster - with booster for COVID-19 detection without RNA purification in less than 1 hour! Our Genaxx1Step RT-qPCR kit for COVID-19 detection is convincing: 1. Kit suitable for gargle testsabout 90% of the test persons prefer a gargle test,if they were able to chose between gargles and sticks 2. Low priceno RNA purification necessary 3. Time-savingless staff involvement required Intended Usage The Genaxx1Step RT-qPCR SARS-CoV-2 Kit  is intended for the use in real-time RT-PCR assays for the in vitro qualitative detection of SARS-CoV-2 Corona virus nucleocapsid fragment target N1 and the control target RNASeP according to the US Centers for Disease Control and Prevention (CDC), which published a protocol for the detection of the 2019-Novel Coronavirus (2019-nCoV). The 2019-nCoV primer and probe sets are designed for the specific detection of 2019-nCoV. Genaxx1Step RT-qPCR SARS-CoV-2 Probe Assay Kit Detection systemGenaxx1Step RT-qPCR SARS-CoV-2 Gargle Kit is a real-time RT-PCR based detection system for the 2019 Wuhan coronavirus (2019-nCoV). 2019-nCoV is considered a novel human coronavirus that is genetically distinct from the common human coronaviruses (229E, NL63, OC43, HKU1), which cause seasonal acute respiratory illness. It is also genetically distinct from the two newer human coronaviruses, MERS-CoV and SARS-CoV. Genaxx1Step RT-qPCR SARS-CoV-2 Gargle Kit detects the presence of the nucleocapsid fragment sequences N1 of 2019-nCoV / SARS-CoV-2 and the control target RNASeP even without prior RNA extraction.The test is not affected by following mutations: B.1.1.7 (British/Alpha), B.1.351 (Southafrican/Beta), B.1.1.28 (Brazilian/Gamma), B.1.525, B.1.617 (Indian/Delta), B1.1.529 (South Africa/Omicron). NOTE: Always include positive and negative controls in your experiment to validate your results. Application: surveillance of cohorts of symptom-free individuals by repetitive testing: The samples are taken repeatedly (e.g. once a week) as described above. PCR pre-screening is carried out without RNA isolation and strong positive virus carriers (up to Ct 30) can be detected quickly and reliably. Subsequently, contact persons of identified virus carriers are tested e.g., with this kit but with prior RNA purification. This ensures that infected persons with Ct over 30 are also identified.