FastGene qFYR Real-Time PCR Systems

Precision device for multiple applications

The FastGene^® qFYR and qFYR Plus are some of the highest precision instruments for performing quantitative polymerase chain reaction (qPCR) experiments. qPCR is a well-established method for sensitive detection and quantification of nucleic acids. During a measurement, the target DNA or RNA sequence is amplified, while a cycle-dependent increase of a fluorescent signal is detected in real-time.

The FastGene qFYR 5-channel system (4+1) and qFYR Plus 6-Channel system were developed to meet highest laboratory standards and deliver reliable performance for most real-time qPCR applications:

• Gene expression analysis
• Absolute and relative quantification
• Endpoint qualitative analysis
• Genotyping
• Gene mutation analysis
• Pathogen detection
• GMO detection
• Protein stability screening
• miRNA analysis
• Melting curve analysis

Highly sensitivity optical design
The FastGene qFYR has a unique, patented optical detection system. It combines a high quality PMT (photomultiplier tube) with a Fresnel lens that has a short focal length. The resulting short distance from detector to sample reduces signal loss and cross-talk between samples, and generally improves signal sensitivity.

The powerful user software has it all
The analysis software (FastGene qFYR Analysis Studio) impresses with its particularly simple operation. The menu is clearly structured and intuitively arranged for different experimental setups, and personalized settings which can be easily adjusted. Integrated analysis algorithms allow many steps, such as baseline subtraction or Cq value threshold calculation, absolute and relative quantification, to be performed automatically.

Generate high quality data
The FastGene^® qFYR is ideal for various quantitative real-time PCR applications with its combination of innovative optics and a high-precision thermal block ensure ideal amplification conditions.   For instance, to the right, the FastGene qFYR shows great homogeneity across all wells of a 96-well plate when amplifying a 1 ng plasmid DNA in all 96 wells of a PCR plate. The mean Cq value at a cycle number of 13.89 +/- 0.055 was determined automatically, illustrating highly homogenous amplification results.

Powerful multiplexing
FastGene qFYR can discriminate up to five different targets in a single reaction well.

Multiplex amplification of three gene targets (Actb, Gapdh and B2m) that was carried out from 100 ng to 0.1 ng RNA in a OneStep qPCR with the FastGene Probe OneStep Mix (LS47).

Accurate quantification
The broad dynamic range of the FastGene qFYR ensures a reliable and accurate quantification.

The amplification plot shows the log of change in normalized reporter fluorescence against the cycle number. The amplification took place from plasmid DNA with AMP specific primers in a 10-fold dilution series ranging from 1 ng to 1×10^-6 ng plasmid DNA. The generated standard curve shows 100 % efficiency.

1.3-fold target discrimination
With the FastGene qFYR, smaller concentration differences can be distinguished with high accuracy, underlining the high sensitivity of the device.

The amplification of plasmid DNA was carried out using AMP-specific primers with a 2-fold dilution series starting at 0.1 ng with additional dilutions of 1:1.3, 1:1.4, 1:1.5 and 1:1.6. Concentration differences can be detected up to 1:1.3-fold dilution.

Melt curve analysis
The Precision Melt Analysis Tool is a powerful method for probe-based allelic discrimination. This function is already included in the FastGene^® qFYR analysis software and does not need to purchased additionally.

The difference plot of the high-resolution melting curve allows the discrimination between SNPs extracted from blood samples for brown eyes and two variants of blue eyes.

Gradient qPCR for optimal annealing
The thermal gradient function of the FastGene qFYR can be used to determine the optimal annealing temperature of a specific target.

Plasmid amplification with IC green dye was carried out in a 10-fold dilution series from 1 ng to 1×10^-6 ng. This example of a thermal gradient experiment with 55 °C and 68 °C shows 55 °C to be the optimal annealing temperature with a 100% efficiency.