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QIAGEN Multiplex PCR Kit

The QIAGEN Multiplex PCR Kit is designed for highly specific and sensitive multiplex PCR without the need for time-consuming optimization

Features and benefits
  • No optimization required easy assay development with convenient ready-to-use master mix
  • High specificity and sensitivity stringent hot start with HotStarTaq DNA Polymerase and increased sensitivity with the unique new multiplex PCR buffer
  • Versatile - highly suited for many types of multiplex PCR applications
  • Easy-to-use and cost-effective simple reaction setup for fast and reproducible results
Principle

The QIAGEN Multiplex PCR Kit is the first kit specifically developed for multiplex PCR and provides an easy-to-use master-mix format. QIAGEN Multiplex PCR Master Mix contains pre-optimized concentrations of HotStarTaq DNA Polymerase and MgCl2, plus dNTPs and an innovative PCR buffer specially developed for multiplex PCR. The new PCR buffer contains a balanced combination of salts and additives, which enables comparable efficiencies for annealing and extension of all primers in the reaction (see figure: "Successful 16plex PCR Using the QIAGEN Multiplex PCR Kit"). Nonspecific annealing is minimized, which leads to maximal yields of specific PCR products. The stringent hot start provided by HotStarTaq DNA Polymerase increases multiplex reaction specificity by preventing extension of nonspecifically annealed primers and primer-dimers. Multiplex assays can be set up at room temperature and HotStarTaq DNA Polymerase in the master mix is then activated by a 15-minute incubation at 95C , which can be incorporated into any existing thermal-cycler program.

Successful 16plex PCR Using the QIAGEN Multiplex PCR Kit
Multiplex PCR of 16 targets (99-955 bp)
was carried out for 35 cycles using standard
conditions for the QIAGEN Multiplex PCR Kit
without further optimization or using a variety
of conditions with a hot-start DNA polymerase
from Supplier AII. A Comparison using 2.5 units
per 50 lreaction of the hot-start DNA polymerase
from Supplier AII and with the indicated Mg2+
concentrations. B Comparison using the optimized
Mg2+ concentration (3.5 mM) for the hot-start
DNA polymerase from Supplier AII (part A) and
the indicated amounts of enzyme per 50 l reaction.
M: markers. Procedure

The QIAGEN Multiplex PCR Kit contains a ready-to-use pre-optimized master mix, to minimize pipetting steps and eliminate tedious calculations. Only primers and template need to be added to prepare the final amplification mix. The master mix can be stored at 2-8C allowing even faster setup of multiplex PCR assays.

Applications

The QIAGEN Multiplex PCR Kit is highly suited for many types of multiplex applications, including:

Genotyping Transgenic Mice

Transgenic mice were screened using the QIAGEN Multiplex PCR Kit and sets of 3 primers to distinguish wild-type (wt), heterozygous mutant (ht), and homozygous mutant (hm) mice. A Using a primer set for the recombination activating gene 2 locus. B Using a primer set for the interferon- V gene locus. M: markers. (Data kindly provided by S. zur Lage and S. Weiss, National Research Center for Biotechnology, Braunschweig, Germany.) Microsatellite Analysis with Optimized QIAGEN Protocol

Analysis of microsatellite loci D3S1358, TH01, D21S11, D18S51, and Penta E was carried out using 1 ng of K562 human genomic DNA and fluoresceinlabeled primers. Reactions were analyzed on the ABI PRISM 377 Sequencer. Top: High sensitivity and uniform signal intensity using the QIAGEN Multiplex PCR Kit. Bottom: Results using a hot-start DNA polymerase from Supplier AII.



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