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Custom and library siRNA for efficient gene silencing

bles a highly stringent hot start and allows reaction setup at room temperature without nonspecific artifacts. The newly developed multiplex PCR buffer in the master mix further enhances the sensitivity of the multiplex reaction. This innovative, new formulation was specifically developed and optimized for multiplex PCR. In contrast to conventional PCR reagents, the new buffer contains a balanced combination of salts and additives to ensure comparable efficiencies for annealing and extension of all primers in the reaction (Figure 1).

Successful 16plex PCR Using the QIAGEN Multiplex PCR Kit

Figure 1 Multiplex PCR of 16 targets (99C955 bp) was carried out for 35 cycles using standard conditions (Std) for the QIAGEN Multiplex PCR Kit (Q) without further optimization or using a variety of conditions with a hot-start DNA polymerase from Supplier AII (AII). A Comparison using 2.5 units/50 l reaction 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 and the indicated amounts of enzyme per 50 l reaction. M: markers.

Versatile for many applications

The QIAGEN Multiplex PCR Kit is well suited for many types of multiplex PCR applications using different detection methods. PCR products can be visualized on agarose gels or detected using fluorescently labeled primers. Detection of fluorescently labeled products can be performed on automatic gel-based DNA sequencers (e.g., the ABI PRISM 377 DNA Sequencer; Figure 2), as well as those based on capillary electrophoresis (e.g., the ABI PRISM 310 Genetic Analyzer).
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