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Real-Time PCR

RealQ Plus 2x Master Mix for Probe

RealQ Plus 2x Master Mix for Probe enables real-time-based DNA amplification with high specificity and efficiency. Real-time PCR is a sensitive and reliable method for gene analysis and DNA quantification. RealQ Plus 2x Master Mix for Probe is the right choice, when expenses and experiment preparation time might be limited or if you need to quickly analyse many genes.

FEATURES

Promotes probe-based real-time PCR based detection
Enables multiplexing
Very high specificity
High stability and efficiency
Reliable quantification
Reaction set-up at room temperature
Premixed all-in-one 2x solution

DESCRIPTION

RealQ Plus 2x Master Mix for Probe is composed of TEMPase Hot Start DNA Polymerase, dNTPs, dye and an optimised buffer system. Furthermore, this master mix is available with high, low or without ROX™ensuring optimal performance on most of the commonly used real-time PCR instruments. Just add DNA template, primers and probe.

The RealQ Plus 2x Master Mix for probe is optimised to TaqMan probes, but is also suitable with other probe chemistries such as Molecular Beacon and Scorpion.

Most probe-based detection methods take advantage of fluorescent resonance energy transfer (FRET) by quenching the signal of a fluorescent reporter in the absence of the desired target. During elongation, the quenching molecule is separated from the fluorescent reporter and a signal is emitted and monitored. Probe-based detection is significantly more specific since signal is only detected when the correct target is amplified.

SELECT THE CORRECT ROX LEVEL

Use the Real-time PCR instrument guide to find the correct level of ROX™ reference dye matching your Real-time PCR instrument.

FAST REAL-TIME PROTOCOLS

The RealQ Plus master mixes have been tested with fast protocols with run time < 50 minutes. The application note can be found here.

HIGH EFFICIENCY AND PRECISION

Amplification plot of a 4-fold dilution series for a Pthr target (75 bp) amplified from human gDNA, starting with 80 ng down to 80 pg of gDNA, using RealQ Plus 2x Master Mix for Probe, High ROX™. Samples were made in triplicates. The results depicted here, show high precision and efficiencies close to 100 %.

REPRODUCIBILITY

80 replicates of RealQ Plus 2x Master Mix Green, High ROX™ and 20 ng gDNA, show a standard deviation of only 0.084.

MULTIPLEXING

Quadruplex real-time PCR reactions. The Pthr1 (FAM), DMD4 (HEX), DMD6 (Texas red) and DMD47 (Cy5) assays are shown in purple, green, pink and orange respectively.

High efficiency, high sensitivity multiplex real-time PCR results with RealQ Flex for Probe. Serial dilution of four different human gDNA targets were amplified either a single-plex real-time PCR, or a 4-target multiplexed real-time PCR.

STABILITY AT ROOM TEMPERATURE

Two plates were pre-assembled for qPCR reaction and incubated in darkness at room temperature for 48 and 72 hours. The results show high stability and complete inactivation of the TEMPase before hot start. It allows the scientist to set up the reaction and run the plate several hours later, when convenient.