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Type-it Microsatellite PCR Kit

For fast and reliable multiplex PCR analysis of microsatellite loci

Features

  • Reliable microsatellite analysis by multiplex PCR
  • Microsatellite assay development without optimization
  • Successful and specific coamplification of all fragments
  • Optimized protocol for fast and reliable results
  • Easy instructions for use with various downstream analysis platforms
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Type-it Microsatellite PCR Kit (200)

Cat. No. / ID: 206243

For 200 x 25 μl reactions: Type-it Multiplex PCR Master Mix (3 x 0.85 ml), 5x Q-Solution (1 x 2 ml), and RNAse-Free Water (2 x 1.9 ml)
$261.00
Reactions
200
2000
Add to cart
The Type-it Microsatellite 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 Microsatellite PCR Kit is based on highly specific HotStarTaq Plus DNA Polymerase and a patented buffer system, both of which enable reliable multiplex PCR-based microsatellite analysis without optimization. The combination of all components provided in the master mix and the specialized formulation result in highly specific amplification of all loci in parallel. The kit is dedicated to fast and reliable microsatellite analysis by multiplex PCR. Optimized protocols are also provided to enable subsequent analysis either by high-resolution capillary sequencing or by using other electrophoresis instruments.

Performance

The Type-it Microsatellite PCR Kit outperformed kits tested from other suppliers and ensures reliable microsatellite analysis. The kit is specifically developed and functionally validated for multiplex PCR-based analysis of microsatellites or minisatellites such as STRs or VNTRs used, for example, in relationship analysis or population genetics (see figure " Reliable 13-plex STR analysis without the need for optimization").
See figures

Principle

The Type-it Microsatellite PCR Kit provides fast and reliable genotyping analysis of humans, animals, plants, and bacteria using microsatellites, STR, or VNTR markers, without the need for lengthy optimization procedures. The Type-it Microsatellite PCR Kit is based on proprietary QIAGEN multiplex technology and includes highly specific HotStarTaq Plus DNA Polymerase, a chemically modified hot-start enzyme, and a specially developed multiplex buffer system including Factor MP. Together, these components ensure high yields and enable reliable multiplex amplification of all microsatellite loci of interest. The kit includes dedicated protocols for the amplification of microsatellites or STRs, as well as step-by-step advice on template amounts, cycle numbers, and PCR conditions and instrument details for different downstream analysis platforms such as agarose gels, capillary sequencers, the Agilent Bioanalyzer, and the QIAxcel Advanced System. 

Type-it Microsatellite PCR Buffer

Unique Type-it Microsatellite PCR Buffer facilitates the amplification of multiple PCR products. In contrast to conventional PCR reagents, the Type-it Microsatellite PCR Buffer contains a specially developed, balanced combination of salts and additives to ensure comparable efficiencies for annealing and extension of all primers in the reaction. Owing to a uniquely balanced combination of KCl and (NH4)2SO4, the PCR buffer provides stringent primer-annealing conditions over a wider range of temperatures and Mg2+ concentrations than conventional PCR buffers. The need to optimize PCR by varying the annealing temperature or the Mg2+ concentration is dramatically reduced, or often not required. Commonly employed optimization procedures for multiplex PCR are virtually eliminated. The buffer also contains the synthetic Factor MP (see figure " Stable and efficient annealing"), which allows efficient primer annealing and extension, irrespective of primer sequence. Factor MP increases the local concentration of primers at the DNA template and stabilizes specifically bound primers.

Q-Solution

The Type-it Microsatellite PCR Kit includes Q-Solution. This innovative PCR additive facilitates amplification of difficult templates by modifying the melting behavior of DNA. Use of this unique reagent will often enable or improve suboptimal PCR. Unlike DMSO and other PCR additives, Q-Solution is used at a defined working concentration with any primer-template system and is not toxic.

See figures

Procedure

Dedicated, application-specific protocols, optimized for analysis of microsatellites on high-resolution capillary sequencers, are included with the Type-it Microsatellite PCR Kit to ensure reliable results for routine analysis, as well as for the establishment of new assays. Reactions can be set up at room temperature, ensuring greater convenience and ease of use.

Fast and simple way to reliable genotyping results

The Type-it Microsatellite PCR Kit enables fast and easy multiplex assay development for accurate and reproducible genotyping results. Whatever the application — microsatellite, STR, SSR, or VNTR analysis — just add the template and primers, and start the thermal-cycler program according to the optimized protocol. The reaction mixture contains all of the reagents required for microsatellite-specific multiplex PCR and enables accurate results without the need for lengthy optimization procedures compared to current methods (see figure " Successful multiplex PCR-based genotyping analysis").

Optimized for use with capillary sequencers

Microsatellite analysis on high-resolution detection systems such as capillary sequencers is often associated with the problem of uneven product yield and huge intensity differences of fluorescent signals. The Type-it Microsatellite PCR Kit overcomes this limitation and ensures high product yields for all amplicons in a multiplex experiment. The kit includes optimized protocols for use with fluorescent primers and outperforms enzyme solutions from other suppliers (see figure " Reliable 13-plex STR analysis without the need for optimization").

See figures

Applications

The Type-it Microsatellite PCR Kit is used to analyze microsatellites, STRs (short tandem repeats), SSRs (simple sequence repeats), and VNTRs (variable number of tandem repeats) and can be used in diverse research fields such as:

  • Species or individual identification    
  • Analysis of relationship
  • Microsatellite instability
  • Population genetics

Supporting data and figures

Specifications

FeaturesSpecifications
ApplicationsMultiplex PCR, Microsatellites, STRs, VNTRs, SSRs
Reaction typePCR amplification
Product useFunctionally validated and developed for reliable Microsatellite Analysis
MastermixYes
Real-time or endpointEndpoint
Sample/target typeGenomic DNA
Single or multiplexMultiplex
With/without hotstartWith

Resources

Brochures & Guides (3)
Second edition — innovative tools
Addressing critical factors and new solutions
Kit Handbooks (1)
For optimization-free and reliable multiplex PCR based analysis of microsatellites
Safety Data Sheets (1)
Download Safety Data Sheets for QIAGEN product components.
Quick-Start Protocols (1)

FAQ

Why is CoralLoad included in the Type-it Mutation Detect, but not in the Type-it Microsatellite PCR Kit?

CoralLoad Dye unfortunately interferes with Capillary Sequencers and increases the risk of damaging the capillaries of these detection platforms. It is not included in the Type-it Microsatellite PCR Kit, since microsatellite loci are analyzed predominantly on Capillary Sequencers due to their high-resolution capability.

By comparison, mutations are very often analyzed on agarose gels, or the QIAxcel System, compatible with CoralLoad dye included in the Type-it Mutation Detect PCR Kit.

 

FAQ ID -2068
Why is maximum amplicon size for the Type-it Microsatellite PCR Kit limited to 500 bp?

The Type-it Microsatellite PCR Kit was optimzed and validated for this size range since microsatellite analysis typically only uses fragments of up to 500 bp in length.

 

 

 

FAQ ID -2060
Which capillary sequencers can be used for detection of amplicons from the Type-it Microsatellite PCR Kits?

All capillary sequencers can be used for detection of PCR product amplified with the Type-it Microsatellite PCR Kit.

 

 

FAQ ID -2063
Can the Type-it Microsaltellite PCR Kit be used on cyclers with fast ramping rates?

Yes. We tested the Type-it Microsaltellite PCR Kit on different commonly used fast and normal PCR cyclers. No significant differences in performance were observed with the systems tested.

 

 

 

FAQ ID -2062
What is the advantage of using the Type-it Microsatellite PCR Kit?

Microsatellite analysis on high resolution detection systems such as capillary sequencers is often associated with the problem of uneven product yield and huge intensity differences of fluorescent signals. The Type-it Microsatellite PCR technology was specifically developed and validated to overcome this limitation and ensure high product yields for all amplicons up to 500 bp long in a multiplex experiment. The kit includes optimized protocols for use with fluorescent primers and outperforms enzyme solutions from other suppliers.

FAQ ID -2059
Does the Type-it Microsatellite PCR Kit always require an annealing temperature of 60°C, or can pre-optimized annealing temperatures for target-primer systems be used?

For established systems, previously optimized annealing temperatures can be used with the Type-it Microsatellite PCR Kit. For new PCR systems, follow the guidelines in Appendix B, 'Design of Multiplex Primers', in the Type-it Microsatellite PCR Handbook.

 

 

FAQ ID -2061