Qiagen, 204654, QuantiFast Multiplex PCR Kit (400)

IMPORTANT NOTE: As announced earlier, the production of the QuantiFast kits has been discontinued since mid-2021. Hence, these products will be available only until stocks last.

For 400 x 25 µl reactions: 3 x 1.7 ml 2x QuantiFast Multiplex PCR Master Mix (with ROX dye), 2 x 2 ml RNase-Free Water

Features

- Sensitive detection of up to 4 targets in 1 tube
- Faster results with time savings of up to 50%
- Successful multiplex PCR without the need for optimization
- Reliable quantification of low- and high-abundance targets

Product Details

QuantiFast Multiplex PCR Kits enable fast and reliable quantification of up to 4 cDNA or gDNA targets in a single tube by multiplex, real-time PCR or two-step RT-PCR. Q-bond technology and an optimized master mix promote fast multiplex real-time PCR, not only on fast cyclers with short ramping times, but also on standard cyclers. The combination of a hot start and a unique PCR buffer system in the ready-to-use master mix ensures highly sensitive qPCR on any real-time cycler without the need for optimization. Two kit formats are available: the QuantiFast Multiplex PCR Kit for cyclers that require ROX dye for fluorescence normalization, and the QuantiFast Multiplex PCR +R Kit for all other cyclers. For convenience, the master mix can be stored at 2–8°C.

IMPORTANT NOTE: As announced earlier, the production of the QuantiFast kits has been discontinued since mid-2021. Hence, these products will be available only until stocks last. Visit the product page of the successor kit to view improved features or to request a trial kit.

For more information and FAQs on this transition, visit: www.qiagen.com/PCRresource .

Performance

The special master mix supplied with QuantiFast Multiplex PCR Kits allows rapid setup of multiplex reactions and delivers successful results at the first attempt, providing multiplex PCR data that are comparable with singleplex PCR data (see figure " Comparable results in triplex and singleplex PCR "). The kits can clearly distinguish between small differences in the amount of template. Even with twofold differences in template amount, QuantiFast Multiplex PCR Kits provide accurate quantification of targets of widely differing abundance (see figure " Linear C T values over twofold decreases in template ").

QuantiFast Multiplex PCR Kits reduce PCR run times by up to 50%, allowing you to get results significantly faster (see figure " Significantly reduced PCR times "). As little as 10 copies of target can be detected in just 60 minutes (see figure " Sensitive duplex PCR and a wide dynamic range "). You can also greatly increase your sample throughput or efficiently share a cycler with other users. Fast results in multiplex, real-time PCR of up to 4 targets are achieved without compromising performance (see figure " Uncompromised sensitivity ").

Principle

Component: Features
HotStarTaq Plus DNA Polymerase: 5 min activation at 95ºC
QuantiFast Multiplex PCR Buffer: Balanced combination of NH 4 + and K + ions
Synthetic Factor MP: Reliable multiplexing analysis of up to 4 genes in the same tube
Unique Q-Bond additive: Faster PCR run times, enabling faster results and more reactions per day
ROX dye †: Normalizes fluorescent signals on Applied Biosystems and, optionally, Agilent instruments

QuantiFast Multiplex PCR Kits deliver highly sensitive and rapid results over a wide dynamic range on both standard and fast cyclers without optimization (see flowchart " QIAGEN multiplex kits "). The specially developed fast PCR buffer contains the novel additive Q-Bond, which significantly reduces denaturation, annealing, and extension times (see figure " Fast primer annealing ").

Amplifying reference and target genes in the same reaction instead of in separate reactions increases the reliability of gene quantification by minimizing handling errors. The QuantiFast Multiplex PCR Buffer includes a balanced combination of K + and NH 4 + ions as well as the unique synthetic Factor MP, which together promote stable and efficient annealing of primers and probes to the nucleic acid template, enabling high PCR efficiency (see figure " Unique PCR buffer "). In addition, HotStarTaq Plus DNA Polymerase provides a stringent hot start, preventing the formation of nonspecific products.

Procedure

ROX dye: Kit
Supplied in master mix: QuantiFast Multiplex PCR Kit
Supplied in separate tube: QuantiFast Multiplex PCR +R Kit

QuantiFast Multiplex PCR Kits contain ready-to-use master mixes that eliminate the need for optimization of reaction and cycling conditions. Simply add template DNA and primer-probe sets to the master mix and follow the protocol in the handbook to get fast and reliable results on any real-time cycler. Kits are available with or without ROX passive reference dye in the master mix, enabling use on virtually any real-time cycler (see table). Due to the optimized ROX concentrations, detection of even low copy numbers is achieved through automatic data analysis.

For optimal results in real-time two-step RT-PCR, we recommend synthesizing cDNA using the QuantiTect Reverse Transcription Kit, which provides fast cDNA synthesis in just 20 minutes with integrated removal of genomic DNA contamination.

QuantiFast Probe Assays are predesigned, genomewide assays that use hydrolysis, probe-based detection. They are delivered with the QuantiFast Multiplex PCR Kit for guaranteed results in duplex, two-step real-time RT-PCR.

Applications

QuantiFast Multiplex PCR Kits can be used for multiplex gene expression analysis of cDNA or gDNA targets on any real-time cycler. This includes instruments from Agilent, Applied Biosystems, Bio-Rad, Cepheid, Eppendorf, and Roche. For the Rotor-Gene Q and other Rotor-Gene cyclers, we recommend using the Rotor-Gene Multiplex PCR Kit, which has been specially developed for fast cycling on these instruments.