Rotor-Gene 3000 Four-Channel Multiplexing System

The Rotor-Gene multi-filter system can detect all available real-time chemistries including Sybr-Green, dual-labelled and MGB probes, FRET and Molecular Beacons.
Most DNA amplification enzymes/ buffers can be used on the system to generate Quantitation/Melt data.

The worlds first centrifugal, real-time DNA amplification system. The fluorescence of up to four different probes can be detected in a single tube. By exciting each dye at its peak wavelength, sensitivity is maximised and cross-talk between channels is minimised.

Using dual labelled probes in a multiplex reaction, genotype data can be generated across a range of loci and disease states. Probes are designed to hybridise specifically to wild type and mutation sequences. A threshold value is set to determine positive or negative amplification so genotypes can be called automatically. S

Blue, green, orange and red are used to evenly span the visible spectrum. Six separate detection filters are selectable providing the standard four channels for multiplexing and an additional two filters for specialised applications. When multiplexing four channels, less than 1% cross-talk is seen between channels and no software algorithm is required to analyse data.

As the samples are spinning at 500rpm there is absolutely no variation in the temperature from sample to sample, a key factor in the precision of this real-time cycling system. This allows for extremely short hold times since no temperature equilibration time is required as with a 96 well block system. Variation in amplification efficiency due to temperature nonuniformity is therefore eliminated

Faster Run Times

By heating the samples only the required amount to initiate denaturation, time is not wasted holding unnecessarily high temperatures. Run times can be reduced by upto 25%.

Optical Denaturation™, by virtue of optical feedback, automatically compensates for any volume being run. Optical Denaturation™ is a patented method that has been developed to take full advantage of the unique features of the Rotor-Gene 3000.

The unit is designed to take data at high speed. All 72 samples can be detected in one revolution, equivalent to 0.15 seconds.

0.01c Temperature Uniformity

All samples cycle without any temperature difference, when one sample denatures, all samples denature. Optical Denaturation™ actively monitors a DNA reference tube during amplification. A typical 2 step amplification requires only the annealing temperature to be defined. During optical denaturation the heating element is activated at full power and the reference sample is monitored at high speed; when the fluorescent level of the sample drops all the samples are denatured and the chamber is cooled back to the annealing temperature.

Quantification

Reproducibility
Due to the optic and thermal design of the system, there is no need to use an internal passive reference.

Sensitivity
The system uses a photo-multiplier (PMT) that can detect a single photon of light. This gives excellent sensitivity even when amplifying a single copy of DNA.

Linearity
The system uses a 16 bit analog to digital converter which has a broad dynamic range.
This results in linear quantization over a wide range of sample concentrations.

Precision
Simple optic design, together with ultimate temperature uniformity (due to centrifugal design) results in a highly accurate and reproducible system.

Our lab uses this machine for the following tests>>

1. Qualitative/Quantitative analysis of MTB
2. Qualitative/Quantitative analysis of HBV
3. Qualitative/Quantitative analysis of HCV
4. Qualitative/Quantitative analysis of HIV