Reverse transcription and qPCR with SYBR® Green .

Our primaQUANT 1STEP CYBR RT-qPCR Master Mix for SYBR® Green RT-qPCR combines cDNA synthesis with quantitative PCR. The product contains all enzymes for both reverse transcription and qPCR, including dNTPs and MgCl₂. You only need to add primers and template RNA. The optimized buffer system ensures fast kinetics and targeted amplification, especially for difficult templates.

Hard Facts .

Applications .

Gene expression / RNA expression in one step

The primaQUANT 1 STEP RT-qPCR Master Mix allows reverse transcription and qPCR in one step. Thus, gene expression can be determined quickly via the RNA level without additional effort.

Pathogen detection

For the diagnosis of pathogenic RNA viruses, the pathogen RNA must be transcribed into cDNA. With the primaQUANT 1Step RT-qPCR Master Mix, you can complete reverse transcription and qPCR in one go. 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 .

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.

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.

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 .

Made in Germany .

We develop and produce our enzymes and master mixes exclusively ourselves in Germany. You benefit from this in several ways:

primaQUANT RT-qPCR Master Mixes with SYBR® Green .

Available with ROX or without ROX – suitable for all qPCR instruments.

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Downloads .

Manuals

Flyer

ROX Compatibility List

Protocol

Manuals

Flyer

ROX Compatibility List

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Hersteller	qPCR Gerät	ROX notwendig?
Applied Biosystems / Thermo Fischer	StepOne™ Real-Time PCR System	Ja (high ROX)
	StepOnePlus™ Real-Time PCR System	Ja (high ROX)
	7500 Real-Time PCR System	Ja
	7500 Fast Real-Time PCR System	Ja
	7500 Fast Dx Real-Time PCR Instrument	Ja
	7500 Real-Time PCR System for Human Identification	Ja
	7300 Real-Time PCR System	Ja (high ROX)
	Viia™ 7 Real-Time PCR System	Ja
	7900HT Fast Real-Time PCR System	Ja (high ROX)
	OpenArray® Real-Time PCR Platform	Ja (high ROX)
	PRISM® 7000 Sequencing Detection System	Ja (high ROX)
	PRISM® 7700 Sequencing Detection System	Ja (high ROX)
	PRISM® 7900 Sequencing Detection System	Ja (high ROX)
	Gene Amp 5700	Ja (high ROX)
	Quantstudio™ 3 / 3 FAST	Ja
	Quantstudio™ 5 / 5 FAST	Ja
	Quantstudio™ 6	Ja
	Quantstudio™ 7	Ja
	Quantstudio™ 12	Ja
	QuantStudio™ 12K Flex system	Ja
Bioneer	Exicycler™96	Nein (optional)
	Exicycler™384	Nein (optional)
Biorad	CFX96™ Real-Time PCR Detection System	Nein
	CFX96 Touch™ Real-Time PCR Detection System	Nein
	CFX384 Touch™ Real-Time PCR Detection System	Nein
	CFX Opus 96 Real-Time PCR System	Nein
	CFX Opus 384 Real-Time PCR System	Nein
	CFX Connect™ Real-Time PCR Detection System	Nein
	iQ5 Real-Time PCR Detection System	Nein
	MiniOpticon™ Real-Time PCR Detection System	Nein
	Opticon 2 – Continuous Fluorescence Detection System	Nein
	Chromo4™ Four-Color Real-Time Detector	Nein
Bioron	RealLine Cycler 48-4	Nein (optional)
	RealLine Cycler 48-5	Nein (optional)
	RealLine Cycler 96-4	Nein (optional)
	RealLine Cycler 96-5	Nein (optional)
DNA Technology	Dtlite	Nein (optional)
	Dtprime	Nein (optional)
Eppendorf	Mastercycler® ep realplex 4, Real-Time Thermal Cycler	Nein
	Mastercycler® ep realplex 4s, Real-Time Thermal Cycler	Nein
	Mastercycler® ep realplex, Real-Time Thermal Cycler	Nein
	Mastercycler® ep realplex s, Real-Time Thermal Cycler	Nein
	Mastercycler Nexus	Nein
	Mastercycler Nexus gradient	Nein
	Mastercycler Pro	Nein
	Mastercycler Pro S	Nein
	Mastercycler Pro 384	Nein
Roche	LightCycler® 96 System	Nein
	LightCycler® 480 System	Nein
	LightCycler® 2.0 Instrument	Nein
	LightCycler® 1.5 Instrument	Nein
	LightCycler® 1536 System	Nein
	LightCycler® Nano System	Nein
Analytik Jena	qTower	Ja
	qTower 2.0	Ja
	qTower 2.2	Ja
	qTower 3	Ja
	qTower 3 84	Ja
Stratagene / Agilent	AriaMx Real-time PCR System	Nein
	Mx3000P® qPCR System	Ja
	Mx3005P® qPCR System	Ja
	Mx4000® qPCR System	Ja
Qiagen	Rotor-Gene™ Q	Ja
	Rotor-Gene™ 6000	Ja
Quantabio	Q	Nein (optional)

Protocol

Composition

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Komponente	Stock Konzentration	20 µl Reaktion	10 µl Reaktion	Finale Konzentration
primaQUANT 1STEP Master Mix	2x	10 µl	5 µl	1 x
Forward Primer	8 µM	1 µl	0,5 µl	400 nm (100 – 600 nM empfohlen)
Reverse Primer	8 µM	1 µl	0,5 µl	400 nM (100 – 600 nM empfohlen)
Probe / Sonde	8 µM	1 µl	0,5 µl	400 nM (100 – 600 nM empfohlen)
Template RNA	10 ng/µl	2 µl	1 µl	0,1 – 100 ng/Reaktion
Steriles Wasser	–	5 µl	2,5 µl	–

Standard protocol

3-Step

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	Schritt	Zeit	Temperatur
Reverse Transkription		10 Minuten	50°C
Initiale Denaturierung		3 Minuten	92°C – 95°C
25 – 45 Zyklen	Denaturierung	5 Sekunden	92°C – 95°C
	Annealing	5 Sekunden	60°C – abhängig von den verwendeten Primern
	Extension	5-10 Sekunden	72°C

2-Step

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	Schritt	Zeit	Temperatur
Reverse Transkription		10 Minuten	50 °C
Initiale Denaturierung		3 Minuten	92 °C – 95°C
25 – 45 Zyklen	Denaturierung	5 Sekunden	92 °C – 95°C
	Kombiniertes Annealing/Extension	10 Sekunden	60 °C – abhängig von den verwendeten Primern

Ultra-FAST protocol

3-Step

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	Schritt	Zeit	Temperatur
Reverse Transkription		5 Minuten	50°C
Initiale Denaturierung		60 Sekunden	92°C – 95°C
25 – 45 Zyklen	Denaturierung	1 Sekunde	92°C – 95°C
	Annealing	1 Sekunde	60°C – abhängig von den verwendeten Primern
	Elongation	1-5 Sekunden	72°C

2-Step

Speichern
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	Schritt	Zeit	Temperatur
Reverse Transkription		5 Minuten	50 °C
Initiale Denaturierung		60 Sekunden	92°C – 95°C
25 – 45 Zyklen	Denaturierung	1 Sekunde	92°C – 95°C
	Kombiniertes Annealing/Elongation	1-5 Sekunden	60°C – abhängig von den verwendeten Primern

Sie haben Fragen?

Steinbrenner Laborsysteme GmbH

Reverse transcription and qPCR with SYBR® Green .

Hard Facts .

Applications .

Features .

For all qPCR cyclers .

RT-qPCR in under 35 minutes .

Direct analysis of RNA with very high sensitivity .

Stable even at room temperature .

Smart Package Size .

Coloring for optimized handling .

Made in Germany .

primaQUANT RT-qPCR Master Mixes with SYBR® Green .

Downloads .

primaQUANT 1STEP CYBR – SL-9551 – Manual 183.46 KB

primaQUANT 1STEP CYBR – SL-9551R – Manual 183.46 KB

primaQUANT 1STEP – RT-qPCR Master Mixe – Flyer 1.64 MB

Composition

Standard protocol

3-Step

2-Step

Ultra-FAST protocol

3-Step

2-Step