Type-it Fast SNP Probe PCR Kit

For accurate and reliable SNP genotyping using TaqMan® or TaqMan® MGB probes

Features

  • Validated using TaqMan® SNP Genotyping Assays
  • Automated allele calling and tight fluorescence clusters
  • Suitable for difficult SNP loci or very low template amounts
  • Up to 40% time savings due to fast cycling procedure
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Type-it Fast SNP Probe PCR Kit (800)

Cat. No. / ID: 206045

For 800 x 25 μl reactions: 6 x 1.7 ml of 2x Type-it Fast SNP Probe PCR Master Mix, 5x Q-Solution, RNase-Free Water
€829.00
Reactions
800
4000
Add to cart
The Type-it Fast SNP Probe PCR Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Product Details

The Type-it Fast SNP Probe PCR Kit is based on highly specific HotStarTaq Plus DNA Polymerase and a newly developed buffer system, both of which enable reliable and clear allelic discrimination. The combination of all components provided in the master mix result in improved accuracy and highly specific probe binding. The Type-it Fast SNP Probe PCR Kit is validated using TaqMan® SNP Genotyping Assays and enables reproducible SNP genotyping — even with difficult SNP loci (e.g., GC rich) or low amounts of starting template.

Performance

The Type-it Fast SNP Probe PCR Kit consistently ensures highly accurate SNP genotyping. 

Highly stringent and specific binding of the allele-specific probe 

The Type-it Fast SNP Genotyping PCR Master Mix is based on highly specific HotStarTaq Plus DNA Polymerase and a newly developed SNP genotyping PCR buffer system, both of which enable highly specific probe binding and consistently strong fluorescent signals. Compared to other commercially available SNP genotyping master mix chemistries, wider and clearer separation of allele clusters is obtained (see figure " Highly specific probe binding").

High call rates even for low template amounts

Even with challenging targets or difficult SNP loci, the Type-it Fast SNP Probe PCR Kit provides reliable SNP genotyping — well-separated allele clusters and tight clustering of identical alleles are achieved, resulting in high call rates  (see figure " Successful automated allele calling"). The Type-it Fast SNP Probe PCR Kit outperformed kits tested from other suppliers, ensuring tight clustering of alleles — even with 1 ng of template DNA (see figure " Reliable SNP genotyping with small amounts of template").

See figures

Principle

The Type-it Fast SNP Probe PCR Kit is available in a convenient master mix format consisting of highly specific HotStarTaq Plus Polymerase and a unique SNP genotyping buffer system designed for fast and efficient amplification. The unique kit components provide tight allele clustering and outstanding separation to ensure high call rates, enabling reproducible and accurate results (see table).

High specificity and sensitivity

HotStarTaq Plus DNA Polymerase, provided in the master mix, which ensures highly specific amplification, even with low template amounts. HotStarTaq Plus DNA Polymerase is provided in an inactive state with no polymerase activity at ambient temperatures. This prevents the formation of misprimed products and primer dimers at low temperatures. 

Unique buffer system

The Type-it Fast SNP PCR Buffer, also included in the master mix, is specifically designed for fast-cycling SNP genotyping using sequence specific 5'-nuclease probes. The unique composition of the Type-it Fast SNP PCR Buffer ensures highly stringent and specific binding of the allele-specific probe (match probe). This is due to the altered melting behaviour of the probes resulting in a narrower probe melting temperature window. Based on the original QIAGEN PCR Buffer, this innovative formulation enables a high ratio of specific-to-nonspecific primer binding during the short annealing step of every PCR cycle. Owing to a uniquely balanced combination of KCl and (NH4)2SO4, the PCR buffer provides stringent primer annealing conditions over a wider range of annealing temperatures and Mg2+ concentrations than conventional PCR buffers. Optimization of PCR by varying the annealing temperature or the Mg2+ concentration is dramatically reduced and often not required.

Additives in the Type-it Fast SNP PCR buffer, such as Q-Solution, provide reaction conditions for amplification of difficult genomic regions and difficult SNP loci. Innovative Q-Bond technology (for fast cycling) allows SNP genotyping results to be achieved faster (see figure " Mechanism of fast cycling during annealing"). The master mix also contains ROX dye at a concentration compatible with SNP genotyping on all instruments from Applied Biosystems. However, ROX dye is also compatible with other instruments not requiring ROX as a passive reference dye (see table). 

Kit features
Kit contents Features
2x Master Mix format* Developed for SNP genotyping with TaqMan® MGB probes
Suitable for use with all cyclers suitable for SNP genotyping
HotStarTaq Plus DNA Polymerase Fast and easy reaction setup at room temperature
Highly specific amplification, even with low template amounts
Type-it Fast SNP Probe PCR Buffer Wide separation of genotype clusters
Tight allele clusters and high allele calling rates
Increased specificity in probe binding
Fast cycling due to Q-Bond Molecule
Q-Solution Tighter clusters and higher signals in scatter plot analysis for highly GC-rich SNP loci
Can further improve suboptimal allele calling
* Includes ROX reference dye, dNTPs, and MgCl2.
See figures

Procedure

The Type-it Fast SNP Probe PCR Kit is functionally verified with commercially available SNP genotyping assays and is compatible with TaqMan® MGB Probes, as well as user-developed probe-based assays consisting of TaqMan® MGB, TaqMan®, or other dual-labeled probes. The kit includes streamlined, preoptimized protocols for fast and reliable analysis.

Fast-cycling procedure for straightforward results

The Type-it Fast SNP Probe PCR Master Mix provides reaction conditions for fast-cycling PCR using sequence-specific probes. The buffer includes proprietary Q-Bond Molecule, which allows short cycling times on standard cyclers and on fast cyclers with rapid ramping rates. The fast-cycling procedure increases throughput by reducing PCR run times by up to 40% (see figure " Genotyping workflow"). 

Convenient kit format

The kit is provided in a ready-to-use, preoptimized master mix format for greater convenience. Use of a master mix saves time, simplifies handling for reaction setup, and increases reproducibility by eliminating many possible sources of pipetting errors and contamination —  pipetting steps are minimized and tedious calculations are eliminated. HotStarTaq Plus DNA Polymerase (included in the master mix) is activated by a 5-minute, 95°C incubation step, which can easily be incorporated into existing thermal cycling programs. Room-temperature reaction setup using the master mix is fast and easy.

Suitable instruments

The Type-it Fast SNP Probe PCR Kit is optimized for amplification of SNP PCR assays on any suitable standard and fast ramping cycler compatible with optical PCR plates that fit into real-time PCR instruments used for fluorescence plate read analysis (see table).

Suitable instruments
Cyclers Model
Real-time PCR cyclers QIAGEN: Rotor-Gene Q
ABI PRISM 7900 (all series) 
ABI StepOne and StepOne Plus
Applied Biosystems 7500 (all series)
Applied Biosystems ViiA 7
Bio-Rad: iCycler iQ
Bio-Rad: CFX series
Roche: LightCycler 480
Roche: LightCycler Nano
Agilent: Mx3000P and Mx3005P
Standard cyclers All cyclers in standard-ramping modes (e.g., GeneAmp 9700)
All cyclers in fast-ramping modes (e.g., GeneAmp 9800)
See figures

Applications

The Type-it Fast SNP Probe PCR Kit is used to detect SNPs using TaqMan® MGB probes and can be used in various fields of research, including:

  • Typing of disease loci
  • Biomarker discovery

Supporting data and figures

Specifications

FeaturesSpecifications
ApplicationsProbe-based SNP Genotyping
Product useFunctionally validated and developed for reliable SNP Genotyping
MastermixYes
Real-time or endpointBoth
With or without ROXROX included in Master Mix
With/without hotstartWith
Sample/target typeGenomic DNA
Enzyme activity5'-> 3' Exonuclease activity
Sequence specific ProbeTaqMan® Genotyping Assays, TaqMan® or TaqMan® MGB probes
Reaction typePCR amplification

Resources

Brochures & Guides (3)
Second edition — innovative tools
Now with even more applications!
Addressing critical factors and new solutions
Kit Handbooks (1)
Safety Data Sheets (1)
Download Safety Data Sheets for QIAGEN product components.

FAQ

Can other probe technologies besides TaqMan® be used with the Type-it Fast SNP Probe PCR Kit?

The Type-it Fast SNP Probe PCR Kit was developed and validated for use with 5'-3' nuclease (hydrolysis) probes like TaqMan® MGB or standard TaqMan® probes. We have not yet tested other probe technologies with this kit.

See trademarks

FAQ ID -2058
Why is special Type-it Fast SNP Probe PCR chemistry required for TaqMan® SNP Genotyping?

SNP Genotyping using TaqMan® probes is a unique application requiring the development of a unique reaction chemistry. The Type-it Fast SNP Probe PCR Kit allows wide separation of genotype clusters and reliable amplification of any difficult genomic region for high call rates.

See trademarks.

FAQ ID -2057