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QIAGEN Multiplex PCR Plus Kit

QIAGEN Multiplex PCR Plus Kit

For straightforward and successful multiplex PCR in advanced applications

  • Successful multiplex PCR for advanced applications
  • Consistent and reliable results
  • Rapid establishment of multiplex PCR assays without optimization
  • Faster reaction times and greater convenience

The QIAGEN Multiplex PCR Plus Kit is designed for easy and sensitive multiplex PCR without the need for optimization. The kit is based on proprietary multiplex PCR technology and delivers successful results at the first attempt, and with far greater speed and ease than ever before. The kit is provided in an easy-to-use master mix format. High specificity and sensitivity is ensured due to the unique composition of the master mix. The master mix contains an innovative buffer specially developed for multiplex PCR. The kit is also provided with Q-Solution, a unique additive that promotes amplification of difficult-to-amplify targets such as GC-rich regions or templates with a complex secondary structure. Handling convenience and visualization of DNA migration is ensured due to CoralLoad Dye — also provided with the kit.

Principle and procedure

The QIAGEN Multiplex PCR Plus Kit is based on QIAGEN's proprietary multiplex PCR technology and is provided in an easy-to-use master mix format. The Multiplex PCR Master Mix contains preoptimized concentrations of HotStarTaq Plus DNA Polymerase and MgCl2, as well as dNTPs and an innovative PCR buffer specially developed for multiplex PCR. The new PCR buffer contains a balanced combination of salts and additives such as Factor MP, which enables comparable efficiencies for annealing and extension of all primers in the reaction (see figure "Unique buffer composition"). View this animation and discover the benefits of QIAGEN's unique dual-cation PCR buffer (see figure "Animation on QIAGEN's unique PCR buffer system"). Nonspecific annealing is minimized and parallel amplification of all targets is successful — even with very low template amounts (see figure "Successful 16-plex PCR over a wide range of template amounts"). The stringent hot-start mechanism provided by HotStarTaq Plus DNA Polymerase increases multiplex reaction specificity by preventing extension of nonspecifically annealed primers and primer–dimers. Optimization of reaction conditions is not required. Multiplex assays can also be easily set up at room temperature. The kit also includes Q-Solution, a unique additive that promotes amplification of difficult-to-amplify targets such as GC-rich regions or templates with a complex secondary structure. Highly versatile CoralLoad Dye — also provided with the kit — further increases handling convenience by improving pipetting visibility and subsequent gel loading and visualization of DNA migration (see figure "CoralLoad Dye for easy PCR setup and convenient DNA visualization"). All of the kit components are specially developed to ensure maximum ease of use and speed, delivering consistently reliable results (see table "Advantages of the QIAGEN Multiplex PCR Plus Kit "). In contrast to alternative approaches — which fail even after lengthy optimization — the QIAGEN Multiplex PCR Plus Kit enables success in multiplex PCR at the first attempt using a single protocol (see figure "Efficient 19-plex PCR using the QIAGEN Multiplex PCR Plus Kit").

Advantages of the QIAGEN Multiplex PCR Plus Kit

 Kit component

 Advantages

HotStarTaq Plus DNA Polymerase

Highly specific and sensitive amplification
Room-temperature setup
5-minute activation time

 Multiplex PCR Plus Buffer with Factor MP

Amplification of all targets in parallel
No optimization of PCR parameters needed
One single protocol for all multiplex assays
Fast procedure

CoralLoad Dye

Easy visualization during pipetting
Immediate gel loading
Visualization of DNA migration

Q-Solution

Amplification of difficult targets

Critical factors for successful multiplex PCR — obtain consistent and reliable results for your applications

Multiplex PCR saves time and reagents for researchers performing large numbers of PCRs and is widely used in genotyping and DNA or cDNA testing applications in research, forensic, and molecular testing laboratories. Applications include typing and analysis of transgenic organisms and pathogens, amplification and analysis of microsatellites, and detection of regions for SNPs or mutations, as well as metagenomics studies (see table "Applications of multiplex PCR"). However, multiplex PCR in general is a highly demanding technique. Factors influencing the success or failure of multiplex PCR are mainly the establishment of optimal PCR parameters. The varying hybridization kinetics of different primer pairs in multiplex PCR can lead to problems such as amplification bias. Primers that bind with high efficiency utilize more of the amplification reaction components, thereby reducing the yield of other PCR products. Due to the larger number of primers, there is also a greater risk of primer–dimer formation and nonspecific priming. Not addressing these challenges leads to poor sensitivity, nonspecific amplification, and biased amplification of selected targets — and therefore inconsistent and unsatisfactory results. Overcoming these bottlenecks requires tedious and time-consuming optimization steps, resulting in increased costs. The QIAGEN Multiplex PCR Plus Kit eliminates these challenges and easily works at the first attempt. The unique composition of the Multiplex PCR Master Mix ensures highly specific and sensitive amplification, even of difficult-to-amplify targets. The kit enables rapid and successful establishment of all multiplex PCR assays, significantly saving time and costs (see figure "Successful multiplex PCR without the need for optimization").

High specificity and sensitivity, without the need for optimization

The preoptimized master mix and unique buffer formulation deliver unparalleled specificity and sensitivity in multiplex PCR applications, eliminating the need for optimization of PCR parameters (see figures "Efficient 19-plex PCR using the QIAGEN Multiplex PCR Plus Kit" and "Successful 16-plex PCR over a wide range of template amounts").

Straightforward and fast downstream analysis

Downstream analysis of multiplex PCR products obtained with the QIAGEN Multiplex PCR Plus Kit is straightforward and fast using a variety of methods. Whether analysis is performed using agarose gels, capillary sequencers, or using the QIAxcel Advanced System (see figure "The QIAxcel Advanced System"), all amplicons can be easily visualized and individual fragments can be reliably distinguished (see figure "Efficient 19-plex PCR using the QIAGEN Multiplex PCR Plus Kit").

Reliable results lead to time and cost savings for advanced applications

The QIAGEN Multiplex PCR Plus Kit is highly suited for a wide variety of advanced applications. The simple reaction setup, fast procedure, and ease of use provided by the kit ensure reproducible results faster, without the need for optimization of PCR parameters. Multiplex assay development is straightforward and easy, ensuring significant time and cost savings in routine research.

Applications

Multiplex PCR saves time and reagents for researchers performing large numbers of PCRs and is widely used in genotyping and DNA or cDNA testing applications in research, forensic, and molecular testing laboratories. The QIAGEN Multiplex PCR Plus Kit can be successfully used for a wide range of applications (see table "Applications of multiplex PCR").

Applications of multiplex PCR

Source of DNA or cDNA

  Application

 

Plants, animals/human

Analysis of satellite DNA (e.g., STR or VNTR analysis)
Typing of transgenic plants/animals
Lineage analysis (e.g., of farm animals)
GMO analysis
Detection of pathogens
Food analysis
Sex determination
Detection of mutations
Amplification of SNP loci
Qualitative and semiquantitative
gene analysis
Splicing isoform identification
Bacteria/viruses Hygiene analysis
Detection of pathogens
Microbial genotyping
Environmental samples Study of metagenomes
Other Pooling of singleplex assays (time and cost savings)
Target enrichment for high-throughput sequencing of ancient DNA(aDNA)

Images

Unique buffer composition.
  Successful 16-plex PCR over a wide range of template amounts.
CoralLoad Dye for easy PCR setup and convenient DNA visualization.
Efficient 19-plex PCR using the QIAGEN Multiplex PCR Plus Kit.
Successful multiplex PCR without the need for optimization.
The QIAxcel Advanced System.
  Animation on QIAGEN's unique PCR buffer system.

 

 
 
 
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