missing translation for 'onlineSavingsMsg'
Learn More
  1. Home
  2. Products
  3. PCR Equipment and Supplies
  4. PCR Supplies
  5. Real Time PCR Products
  6. Real Time PCR Reagents and Kits
  7. SuperScript™ EXPRESS One-Step Superscript™ qRT-PCR Kit, with premixed ROX

SuperScript™ EXPRESS One-Step Superscript™ qRT-PCR Kit, with premixed ROX

Product Code. 10574414
Change view
Click to view available options
No. of Reactions:
2500 Reactions
500 Reactions
Unit Size:
2500 reactions
Each
2 product options available for selection
Product selection table with 2 available options. Use arrow keys to navigate and Enter or Space to select.
Product Code. No. of Reactions unitSize
10574414 500 Reactions Each
10174783 2500 Reactions 2500 reactions
Use arrow keys to navigate between rows. Press Enter or Space to select a product option. 2 options available.
2 options
This item is not returnable. View return policy

For Research Use Only. All usage must comply with product instructions.
Product Code. 10574414 Supplier SuperScript™ Supplier No. 11791200

Please to purchase this item. Need a web account? Register with us today!

This item is not returnable. View return policy

For Research Use Only. All usage must comply with product instructions.

Description

Next-generation, real-time qRT-PCR reagents designed to complement fast-cycling instruments. Get the turnaround time you want with the data quality you need.

EXPRESS One-Step SuperScript™ qRT-PCR SuperMix Kits incorporate proven Invitrogen enzymes now specifically tailored to deliver superior results on high-throughput, fast-cycling instruments for your probe-based qPCR experiments.

  • Fast-activating, antibody-mediated Platinum™ Taq DNA Polymerase - complete, rapid activation provides the best sensitivity and reproducibility
  • The One-Step SuperScript™ III Reverse Transcriptase module has a unique formulation to ensure better yield and less PCR inhibition, specifically for one-step qRT-PCR
  • Flexible formats with ROX premixed or separate - designed to work with default instrument protocols on a wide range of high-throughput fast instruments (e.g., AB 7500 Fast, AB 7900 HT Fast, AB StepOne, Roche LightCycler™ 480, Corbett Rotor-Gene™)
  • The only commercially available one-step qRT-PCR kits containing heat-labile UDG - significantly reduced carryover contamination risk with excellent activity from room temperature to approximately 40°C. Due to the thermostability of SuperScript™ III, the cDNA synthesis step is run at 50°C, at which temperature the heat-labile UDG is totally and irreversibly inactivated

Easy to test and switch:
  • EXPRESS kits work with manufacturer's default protocols on fast instruments and come in a convenient and stable SuperMix format for ultimate convenience
  • The kits are offered with either premixed ROX or a separate tube of ROX to fit your instrument needs

Order Info

Guaranteed to be stable for 6 months when properly stored.

TRUSTED_SUSTAINABILITY

Specifications

Content And Storage EXPRESS One-Step SuperScript™ qRT-PCR SuperMixes are composed of 2 modules: an EXPRESS RT module (containing SuperScript™ III Reverse Transcriptase and RNAseOUT™) and an EXPRESS qPCR SuperMix module with ROX premixed or in a separate tube.

Store at -20°C upon receipt. Once thawed, the EXPRESS qPCR SuperMix can be stored for up to 1 month at +4·C. Product is guaranteed stable for 6 months when properly stored.
Detection Method Primer-probe
Format Tube
PCR Method 1-step RT-qPCR
Polymerase Taq DNA Polymerase
Reaction Speed Fast or Standard
For Use With (Equipment) 7000 System, 7300 System, 7700 System, 7900HT Fast System, 7900HT System, Eppendorf™ MasterCycler™ RealPlex, StepOne™, Fast Mode, StepOne™, Standard Mode
Product Line SuperScript
Product Type qRT-PCR Kit
Quantity 500 reactions
Sufficient For 500 Reactions
For Use With (Application) Gene Expression
No. of Reactions 500 Reactions
Sample Type RNA
Show More Show Less

Frequently Asked Questions (FAQs)

What can I do to improve the sensitivity of my qPCR assay?

If you are targeting a low-abundance gene, you may have trouble getting Ct values in a good, reliable range (Ct > 32). To increase the sensitivity of the assay, you may want to consider the following:

- Increase the amount of RNA input into your reverse transcription reaction, if possible
- Increase the amount of cDNA in your qPCR reaction (20% by volume max)
- Try a different reverse transcription kit, such as our SuperScript VILO Master Mix, for the highest cDNA yield possible
- Consider trying a one-step or Cells-to-CT type workflow (depending on your sample type)

How do I set the baseline for my qPCR experiment?

Most times your instrument software can automatically set a proper baseline for your data. Check out our short video, Understanding Baselines, for more information on how to set them (https://www.youtube.com/watch?feature=player_embedded&v=5BjFAJHW-bE).

How do I set the threshold for my qPCR experiment?

In most cases your instrument software can automatically set a proper threshold for your data. Check out our short video, Understanding Thresholds, for more information on how to set them (https://www.youtube.com/watch?feature=player_embedded&v=H_xsuRQIM9M).

I am not getting any amplification with my TaqMan Assay or SYBR Green primer set. What is causing this?

There could be several reasons for no amplification from an assay or primer set. Please see these examples and suggested solutions in our Real-Time Troubleshooting Tool (https://www.thermofisher.com/us/en/home/life-science/pcr/real-time-pcr/qpcr-education/real-time-pcr-troubleshooting-tool/gene-expression-quantitation-troubleshooting/no-amplification.html) for more details.

I am getting amplification in my no-template control (NTC) wells in my qPCR experiment. What is causing this?

There could be several reasons for amplification in a NTC well. Please see these examples and suggested solutions in our Real-Time Troubleshooting Tool (https://www.thermofisher.com/us/en/home/life-science/pcr/real-time-pcr/qpcr-education/real-time-pcr-troubleshooting-tool/gene-expression-quantitation-troubleshooting/amplification-no-template-control.html) for more details.

My amplification curves have a funny shape in my qPCR experiment. What is causing this?

There are several reasons that amplification could be delayed. Please see the information in our Real-Time Troubleshooting Tool (https://www.thermofisher.com/us/en/home/life-science/pcr/real-time-pcr/qpcr-education/real-time-pcr-troubleshooting-tool/gene-expression-quantitation-troubleshooting/abnormal-amplification-curves/amplification-occurs-later.html) for more details.

What can I do if the amplification of my target gene is later than expected for my qPCR experiment?

There are several reasons that amplification could be delayed. Please see the information in our Real-Time Troubleshooting Tool for more details (https://www.thermofisher.com/us/en/home/life-science/pcr/real-time-pcr/qpcr-education/real-time-pcr-troubleshooting-tool/gene-expression-quantitation-troubleshooting/abnormal-amplification-curves/amplification-occurs-later.html).

Can I use my SYBR Green primers for a TaqMan assay?

It may be possible to use your SYBR Green primers for a TaqMan assay, depending on how they were designed. You would have to design a separate probe to use with your existing primers. Please refer to the guidelines in this manual (https://tools.thermofisher.com/content/sfs/manuals/cms_041902.pdf) on “Manually Designing Primers and Probes” for the next steps. If you have Primer Express Software, you can use that software to design a probe. Please note that restricting the design using the predesigned SYBR primers may not allow for a successful probe design.

Do I have to normalize my samples for comparative Ct experiments?

Comparative Ct experiments use an endogenous control gene to normalize the cDNA input. Please watch this short video (https://www.youtube.com/watch?feature=player_embedded&v=jst-3hD_xFQ) for more details on how this works. For a protocol workflow, please refer to our Guide to Performing Relative Quantitation of Gene Expression (https://tools.thermofisher.com/content/sfs/manuals/cms_042380.pdf).

What are the requirements for a relative quantification qPCR experiment?

In a relative quantification experiment, you will need to identify an endogenous control and a reference (or calibrator) sample. An endogenous control is a gene that does not change in expression across all the samples in your study. A reference sample is the sample that you are comparing all others to. This is often the untreated, or control, sample. Please see our Relative Gene Expression Workflow bulletin (https://tools.thermofisher.com/content/sfs/brochures/cms_075428.pdf) for more step-by-step guidelines on how to design your experiment.

What are the requirements for a standard curve qPCR experiment?

In a standard curve experiment, you must generate a standard curve for each target gene. The standards should closely represent the sample (i.e., RNA for RNA input, plasmid or gDNA for DNA input). This reference (http://www.ncbi.nlm.nih.gov/pubmed/11013345) is a good review of standard curves and the experimental setup. You can also review this short video (https://www.youtube.com/watch?v=mE5ieko9_RQ) on standard curve experiments.

What is the difference between absolute quantification (AQ) and relative quantification (RQ)? How do I choose which method to use?

Absolute quantification will quantitate unknowns based on a known quantity. It involves the creation of a standard curve from a target of known quantity (i.e., copy number). Unknowns can then be compared to the standard curve and a value can be extrapolated. Absolute quantification is useful for quantitating copy number of a certain target in DNA or RNA samples. The result usually is a number followed by a unit, such as copy number and ng, etc.

Relative quantification can quantitate a fold difference between samples. It involves the comparison of one sample to another sample (calibrator) of significance. For example, in a drug treatment study you could compare a treated to an untreated sample. The quantity of the calibrator is not known and cannot be measured absolutely. Therefore the calibrator (untreated sample) and samples (treated samples) are normalized to an endogenous control (a gene that is consistently expressed among the samples) and then compared to each other to get a fold difference. Relative quantification is useful for quantitating messenger RNA levels. Since the result is a fold change or ratio, it is not followed by a unit.

The method that you choose will depend on the type of data you need from your experiment. You can find more information here (https://www.thermofisher.com/us/en/home/life-science/pcr/real-time-pcr/qpcr-education/absolute-vs-relative-quantification-for-qpcr.html) as well.

Can I do a melt curve with a TaqMan assay?

No. A TaqMan probe, once cleaved, cannot be re-quenched. Therefore a melt curve does not apply when using a TaqMan assay.

What is the difference between TaqMan and SYBR Green methods of detection?

TaqMan and SYBR Green chemistries are two different methods of detection for qPCR. Please see this detailed comparison of these two approaches (https://www.thermofisher.com/us/en/home/life-science/pcr/real-time-pcr/qpcr-education/taqman-assays-vs-sybr-green-dye-for-qpcr.html). You can also watch this short video (https://www.youtube.com/watch?feature=player_embedded&v=fkUDu042xic) on how TaqMan assays work.

How many replicates do I need to run for my qPCR experiment? What recommendations do you have for plate setup?

Please view this short video (https://www.youtube.com/watch?v=eIaPGhOjBQo), which explains some best practices for replicates and plate setup.

What are the different phases of a qPCR reaction?

Check out this short video (https://www.youtube.com/watch?feature=player_embedded&v=4sXPUbIrh3A) to understand the different phases of the PCR reaction and why they are important.

I'm trying to decide between purchasing a one-step or two-step RT-PCR kit. Can you review the advantages and disadvantages of each?

One-step RT-PCR is convenient and less prone to contamination, as there is less opportunity for pipetting error. This method is also faster than the two-step process. However, the cDNA cannot be archived, and fewer genes can be analyzed. Two-step RT-PCR gives you the ability to archive cDNA, analyze multiple genes, and offers greater flexibility. Learn more about the difference between one-step and two-step RT-PCR on this page Onestep vs Twostep RT-PCR.

For Research Use Only. Not for use in diagnostic procedures.

Product Title
Select an issue

By clicking Submit, you acknowledge that you may be contacted by Fisher Scientific in regards to the feedback you have provided in this form. We will not share your information for any other purposes. All contact information provided shall also be maintained in accordance with our Privacy Policy.