RT-qPCR with SYBR®Green .

Our primaQUANT 1-STEP RT-qPCR Master Mix with SYBR® Green combines cDNA synthesis with quantitative PCR. All enzymes for both reverse transcription and qPCR are included in the Master Mix, including dNTPs and MgCl₂

Start your qPCR directly with RNA – no separate cDNA synthesis necessary.

All you have to do is add primers, probes and template RNA. The optimized buffer system guarantees fast kinetics and targeted amplification even with difficult templates.

Hard Facts .

Excellent sensitivity < 10 fg RN
cDNA synthesis and qPCR in one reaction
Hot-start polymerase
Ultra-Fast: RT-qPCR in < 35 minutes
Safe handling due to blue coloring
One-tube format - 1 master mix
Developed & manufactured in Germany

Applications .

Gene expression via RNA level

The primaQUANT 1 STEP RT-qPCR Master Mix is ideally suited for the gene expression analysis. The perfectly matched combination of reverse transcriptase and qPCR in this Master Mix allows you to quickly determine gene expression without separate cDNA synthesis.

Pathogen detection

The diagnosis of pathogenic RNA viruses by qPCR requires the transcription of pathogen RNA into cDNA. With the primaQUANT 1Step RT-qPCR Master Mix, you complete cDNA synthesis and qPCR analysis in one step. This makes high-throughput analysis and automation easy.

Melting curve analysis

High Resultion Melting Curve Analysis (HRM) can be used to easily detect the smallest changes in the genome, such as SNP and mutations, using qPCR.

Matsuda, K. (2017). PCR-Based Detection Methods for Single-Nucleotide Polymorphism or Mutation: Real-Time PCR and Its Substantial Contribution Toward Technological Refinement. Advances in Clinical Chemistry, 80, 45-72. https://doi.org/10.1016/bs.acc.2016.11.002

Simko, I. (2016). High-Resolution DNA Melting Analysis in Plant Research. Trends in Plant Science, 21(6), 528-537. https://doi.org/10.1016/j.tplants.2016.01.004

Mohammad Rahimi, H., Pourhosseingholi, M. A., Yadegar, A., Mirjalali, H., & Zali, M. R. (2019). High-resolution melt curve analysis: A real-time based multipurpose approach for diagnosis and epidemiological investigations of parasitic infections. Comparative Immunology, Microbiology and Infectious Diseases, 67, 101364. https://doi.org/10.1016/j.cimid.2019.101364

Features .

RT-qPCR in under 35 minutes .

In the primaQUANT 1STEP RT-qPCR Master Mixes, we combine optimized reverse transcriptases with an extremely fast DNA polymerase. Thus, a complete RT-qPCR (transcription of RNA into cDNA and subsequent qPCR) can be performed in less than 35 minutes, depending on the qPCR instrument.

This saves you valuable time and capacity, especially for diagnostics or larger numbers of samples.

Of course, you can also continue to use your previous RT-qPCR protocol.

Mit dem primaQUANT 1STEP RT-qPCR Mastermix können Sie eine RT-qPCR in weniger als 35 Minuten durchführen.

For all qPCR cyclers .

Our qPCR 2x Master Mixes are suitable for all common qPCR cyclers and optionally available with ROX.

We have compiled a table for you showing whether your device uses ROX(6-carboxy- X-rhodamine) for normalization and which concentration, if any, is required.

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Direct analysis of RNA with very high sensitivity .

The primaQUANT 1STEP RT-qPCR master mixes are very sensitive and show a wide linear dynamic range.

The product is therefore particularly interesting for diagnostics, where speed and sensitivity are especially important.

Stable even at room temperature .

Freeze/thaw

Our primaQUANT 1STEP Master Mix can also withstand multiple freezing/thawing . You can therefore freeze and defrost the mixes more than 10 times without any problems.

Refrigerator storage

The primaQUANT 1 STEP Master Mixes are stable for weeks at 4°C. For short-term storage, we therefore generally recommend storage at 4°C in the refrigerator.

Stability at room temperature

We have optimized our Master Mixes so that they can withstand several days without cooling without damage. Therefore, you no longer need to pipette your RT-qPCR laboriously on ice to avoid loss of sensitivity. This also makes the primaQUANT 1STEP perfect for automation solutions, as no cooling is required.

Der primaQUANT 1STEP RT-qPCR Master Mix ist mehrere Wochen bei 4°C stabil und kann daher für die schnellere Verwendbarkeit im Kühlschrank gelagert werden.

Durch die besondere Formulierung des primaQUANT 1STEP Master Mixes bleibt die Performance auch nach 10 Einfrier/Auftau-Vorgängen stabil.

Smart Package Size .

No more annoying aliquoting!

Our Master Mixes come in units of 1 ml as standard.

This is very convenient and practical, because one tube is enough for exactly one 96 well plate, two tubes for one 384 well plate. If you ever need less, you can simply store the mixes in the refrigerator for several weeks.

Customized fillings: As the manufacturer, we can of course supply you with fillings and mixtures according to your special requirements.

Coloring for optimized handling .

The primaQUANT 1STEP master mix is colored blue for better handling. This makes pipetting much more convenient and also much safer: You can see immediately whether mastermix has already been added to a preparation and if wells have already been pipetted or not. vvvv

The blue dye used has no measurable adverse effect on RT-qPCR .