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 Master Mixes
  7. Applied Biosystems™ TaqMan™ Genotyping Master Mix

Applied Biosystems™ TaqMan™ Genotyping Master Mix

Catalog No. 4381657
Encompass
Change view
Click to view available options
Quantity:
1 x 1 mL
1 x 10 mL
1 x 50 mL
2 x 10 mL
2 x 50 mL
5 product options available for selection
Product selection table with 5 available options. Use arrow keys to navigate and Enter or Space to select.
Catalog No. Quantity
4381657 2 x 50 mL
4371353 1 x 1 mL
4371355 1 x 10 mL
4381656 2 x 10 mL
4371357 1 x 50 mL
Use arrow keys to navigate between rows. Press Enter or Space to select a product option. 5 options available.
5 options
Catalog No. 4381657 Supplier Applied Biosystems™ Supplier No. 4381657

Please call Customer Service at 1-800-234-7437 or send an email to help@thermofisher.com for assistance.

Description

Optimized for end-point fluorescence detection in SNP genotyping applications.

  • Distinct clusters and high call rates for unambiguous allelic discrimination
  • Validated with TaqMan™ SNP Genotyping Assays
  • Excellent pre- and post-PCR stability for high throughput setup and analysis
  • Reaction size is based on a 50μL reaction volume

May be used for a variety of genotyping applications, including:
  • Candidate gene studies
  • Drug-target validation
  • Disease association studies
  • Population genetics
  • Linkage mapping
  • Agricultural applications

Exceptional Performance from an Optimized Mix (2X)
Accurate genotype assignments result from preferential binding of the allele-specific probe to the matching target. TaqMan™ Genotyping Master Mix enables specific binding of the probe to achieve exceptional cluster resolution. Key features include:
  • AmpliTaq Gold™ DNA Polymerase, UP (Ultra Pure) - Automatic hot start enzyme designed to be active during thermal cycling and inactive at room temperature for easy reaction setup
  • Optimized mix components provide excellent specificity for discrimination between alleles
  • Passive internal reference based on proprietary ROX™ dye for increased precision on Applied Biosystems™ real-time PCR instruments
  • Single thermal cycling condition for consistent results with TaqMan™ Assays

Validated with TaqMan™ Genotyping Assays and Instruments:
  • TaqMan™ Genotyping Master Mix has been tested across all types of TaqMan™ SNP Genotyping Assays: TaqMan™ Drug Metabolism Genotyping Assays, TaqMan™ SNP Genotyping Assays, and Custom TaqMan™ SNP Genotyping Assays
  • In addition, the master mix is validated with the Applied Biosystems™ thermal cyclers and real-time PCR systems

High-Throughput Setup and Analysis:
  • The combination of several low-cost thermal cyclers for PCR and a single real-time PCR instrument for allelic discrimination make high throughput SNP genotyping manageable
  • TaqMan™ Genotyping Master Mix further improves high-throughput setup and analysis with excellent room temperature stability both before and after PCR
  • This provides the flexibility required to run long experiments, unattended overnight or over a weekend

Reliable Discrimination with Challenging Targets
  • Amplicon design constraints make SNP detection challenging, particularly when targets contain an abundance of GC- or AT-rich regions
  • The optimized components of TaqMan™ Genotyping Master Mix result in tight, well-separated clusters and more accurate allele-calls compared to other mixes, even for challenging targets

Setting the Industry Standard for Quality:
  • Extensive analytical and functional tests are performed for each manufactured lot of TaqMan™ Genotyping Master Mix, including one TaqMan™ SNP Genotyping Assay and one TaqMan™ Drug Metabolism Genotyping Assay
  • Results are compiled in an informative Certificate of Analysis, which is accessible through the Applied Biosystems™ Web site

Order Info

Guaranteed minimum shelf life is 60 days. Shipping Conditions: Wet Ice

Specifications

Concentration 2X
Content And Storage Supplied at 2X concentration. The mix is optimized for SNP genotyping using TaqMan™ probes and contains AmpliTaq Gold™ DNA Polymerase UP (Ultra Pure), dNTPs without dUTP, Passive Reference 1 and optimized mix components. This pack contains two 50 mL bottles, sufficient for 4,000 reactions at the 50 μL total volume.

Storage: 2-8°C

Guaranteed minimum shelf life is 60 days (exact expiry date printed on product and CofA).
Detection Method Primer-probe
GC-Rich PCR Performance High
PCR Method qPCR
Polymerase AmpliTaq Gold DNA Polymerase
Reaction Speed Standard
Technique SNP Genotyping (Real-Time PCR-Based), CNV (Copy Number Variation)
For Use With (Equipment) Applied Biosystems StepOnePlus™ Fast Real-Time PCR System, QuantStudio™ 12k Flex, QuantStudio™ 3, QuantStudio™ 5, QuantStudio™ 6 Flex, QuantStudio™ 7
Genotyping Target Copy Number Variants, SNPs (Unknown or Numerous), Drug Metabolizing Enzymes, SNPs (Known)
Passive Reference Dye ROX (Pre-mixed)
Product Line TaqMan
Product Type Genotyping Master Mix
Purity or Quality Grade UP (Ultra Pure)
Quantity 2 x 50 mL
For Use With (Application) Genotyping
Fidelity (vs. Taq) 2 X
Sample Type DNA (Genomic)
Volume 100 mL
Show More Show Less

Frequently Asked Questions (FAQs)

What master mix can I use with the TaqMan SNP Genotyping Assays?

We recommend using either the TaqPath ProAmp Master Mix or the TaqMan Genotyping Master Mix. The TaqMan Genotyping Master Mix has the advantage of proven performance with up to 3 days of pre- and post-PCR stability, allowing you to set up plates ahead of time or read the plates later (see the data here, https://tools.thermofisher.com/content/sfs/brochures/cms_039236.pdf) while the TaqPath ProAmp Master Mix can handle samples that may have inhibitors present.

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.
My sample contains inhibitors of PCR. What master mix should I use for my genotyping and copy number assays?

The validated and recommended mastermix for TaqMan SNP Genotyping assays as well as TaqMan Copy Number Variation Assays is the TaqMan Genotyping Master Mix (Cat. No. 4371353). However, in the event that the sample contains inhibitors of PCR such as those from human or animal sources (buccal swabs, blood, and card punches), you may use the TaqPath ProAmp Master Mix (Cat. No. A30866) for SNP Genotyping as well as for Copy Number Variation analysis.
I would like to order a TaqMan SNP Genotyping Assay. Does the kit include everything I need to perform my experiment or do I need to purchase reagents separately?

Each predesigned TaqMan SNP Genotyping Assay is delivered in a single tube consisting of two differentially labeled allele-specific TaqMan MGB (minor groove binding) probes and a pair of PCR primers. For more information, please see the following link: https://www.thermofisher.com/us/en/home/life-science/pcr/real-time-pcr/real-time-pcr-applications/genetic-variation-analysis-using-real-time/snp-genotyping-with-real-time-pcr.html

Besides the TaqMan SNP Genotyping Assay, you would need to purchase a Genotyping Master Mix and nuclease-free water. A list of compatible master mixes can be found in the following link: https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0009593_TaqManSNP_UG.pdf.

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