The RealQ Fast 2x Master Mix Green offers a high-quality, convenient solution for dye-based real-time PCR workflows. With its optimized formulation, this ready-to-use master mix ensures sensitive, reliable, and rapid results, facilitated by the inclusion of 2Taq Hot Start DNA Polymerase.
FEATURES
Room temperature reaction setup
Multiple choices for fast or super-fast PCR programs
High specificity, stability, and reproducibility
DESCRIPTION
The key to RealQ Fast's performance lies in the antibody-mediated hot-start mechanism of 2Taq, providing precise control over enzyme activity. This feature prevents undesirable amplification at low temperatures, minimizing the risk of nonspecific amplification.
For real-time PCR instruments requiring a reference dye such as ROX, the separate ROX solution is available, allowing for adjustment of ROX levels. This adaptability ensures compatibility with a wide range of real-time PCR instruments, making RealQ Fast a versatile choice for researchers.
For detailed guidelines on optimizing ROX levels to your specific real-time PCR instrument, please refer to the documentation available at ampliqon.com.
SENSITIVE DETECTION AND ACCURATE QUANTIFICATION
The RealQ Fast 2x Master Mix Green provides sensitive detection and quantification across a wide range of DNA templates including human genomic DNA, plasmid DNA and bacterial DNA.
The efficiency and detection limits of RealQ Fast 2x Master Mix Green were assessed using qPCR targeting PAH across a 6-log dilution range of human genomic DNA on an ABI StepOnePlus real-time PCR instrument. The obtained results demonstrate an efficiency of 101% and a detection limit of 0.001 ng (equivalent to 0.6 copies of diploid template). These findings attest to the high sensitivity and accuracy of the master mix. Comparable results were observed for other human gene targets as well as plasmid and bacterial targets, further validating the robust performance of RealQ Fast across diverse applications.
ROOM TEMPERATURE STABILITY
The incorporation of an antibody-mediated hot-start mechanism in the RealQ Fast 2x Master Mix enables users to prepare reactions up to 72 hours before PCR cycling, without compromising performance. This feature not only offers convenience but also underscores the stability and reliability of the master mix. RealQ Fast provides users with a high level of flexibility, instilling confidence in its performance across a wide range of workflows.
The robustness of RealQ Fast 2x Master Mix Green was evaluated through qPCR targeting Pthr1 across a 16 – 0.2 ng/µl dilution range of human genomic DNA. In a comparative analysis, two 96-well plates were prepared with reaction mix, including RealQ Fast 2x Master Mix, primers, and DNA template. One plate underwent immediate PCR cycling, while the other was incubated at room temperature for 72 hours before cycling. Both plates maintained consistent efficiency, R2, and curve shape.
REDUCED RUN TIME
RealQ Fast Green 2x Master Mix has been optimized to deliver sensitive detection and accurate quantification even when a low run time is required. RealQ Fast offers a high degree of flexibility in PCR program design, while maintaining efficiency, high specificity, and high sensitivity*.
*For set-ups demanding a low LOD, it is recommended to utilize the Standard or Fast programs.
A 5-log human genomic DNA dilution range was used to assess the effect of decreased cycle time. The human Pthr1 gene was amplified with three different programs (Standard, Fast and Super-Fast) on the ABI StepOnePlus real-time PCR instrument. The three programs yielded similar results, with efficiencies at 95 - 105% and a LOD of 0.01 ng. Notably, the use of Fast program resulted in a delay of 0.5 Cq-values, while the Super-Fast program resulted a delay of 1.5 Cq-values, in comparison to the Standard program.
