Opticon®

Real-Time SystemsOpticon®

Real-Time Systems

DNA Engine DNA Engine

High-Resolution Real-Time in One or Two Colors

*Please note, the information here is being provided for research purposes only. The equipment and the infor-mation presented have not been approved for, and should not be used for, human diagnostic applications.

The Polymerase Chain Reaction (PCR) is a process covered by US Patents #4,683,195 and #4,683,202, whichare owned by Hoffmann-La Roche. Users should obtain a license to perform the reaction, and a license is currentlyavailable through either Roche Molecular Systems of Pleasanton, California; or F. Hoffman-La Roche, Ltd. ofBasel, Switzerland.

1024

512

256128

Max–Min = 0.88 C(t)

SYBR Green

High resolution—two-fold differences in starting copy numberare easily distinguishable. Log–fluorescence vs. cycle number duringamplification of plasmid containing β-actin cDNA. The experiment wasperformed in replicate (n=6). Initial template copy number is indicatedfor each dilution set.

Uniformity—Log-fluorescence plots show the consistency inC(t) values across wells. Plasmid containing β-actin was amplifiedon an Opticon 2 system. (95 replicates; single no-template control notshown.) Initial template copy number =106.

Advanced thermal control & gradient featureThe Opticon systems incorporate the precise thermal control of aDNA Engine cycler. The accurate, uniform thermal control yieldsreliable, reproducible results, and a temperature gradient featureallows you to optimize cycling conditions in a single experiment.

Innovative optical design for extraordinarysensitivity and low maintenanceBoth Opticon systems incorporate a robust optical design with nomoving parts. Ninety-six light-emitting diodes (LEDs) fire in rapidsequence to illuminate individual wells, resulting in minimal crosstalk.Emitted fluorescence passes through filters to photomultiplier tubes(PMTs), the most sensitive optical detectors available for biologicalapplications.

Because LEDs are durable, with a stable output over time, wells canbe illuminated with little variation, experiment after experiment, withlittle maintenance to the optical system. PMTs amplify signal, improv-ing signal-to-noise ratio so small signals—and small differences insignal—can be discerned. This unique design permits reliabledetection of one initial template copy, while delivering a linearrange of up to 10 orders of magnitude in starting copy number.

Intuitive softwareResearcher-oriented software makes experimental setup and analysiseasy. Go from start to run in just three steps; observe data acquisitionin real-time; and begin data analysis while the run is in progress.Systems are pre-calibrated for a broad range of dyes, includingFAM™ and SYBR® Green I on the Opticon system, and additionaldyes and multicolor capability on the Opticon 2 system. Meltingcurve analysis is included for verifying product identity, and a specialanalysis feature allows automated scoring of genotypes.

Performance & application data

FAM detectionof ERBB2

VIC detectionof GAPDH

Carcinoma

Two-color multiplexing simplifies and improves the accuracy of geneexpression analysis. Results from a 2-color, duplex experiment comparing theexpression of the ERBB2 gene in healthy (above) and carcinoma (below) breast tissuesamples. Amplification of ERBB2 and a housekeeping gene (GAPDH) was monitored withdifferently labeled hydrolysis probes. Note that the C(t) of ERBB2 for the carcinoma isabout four cycles earlier, which indicates approximately sixteen-fold greater expressionof this gene.*

FAM detectionof ERBB2

VIC® detectionof GAPDH

Healthy tissue

Real-time detection is a powerful tool that simplifies DNA

quantification, genotyping, and many other applications.

The MJ Research® Opticon™ and Opticon 2 continuous

fluorescence detection systems feature an innovative optical

design and a built-in DNA Engine® thermal cycler for

extraordinary real-time performance. The Opticon systems

produce reliable, high-resolution results, and the

accompanying Opticon Monitor™ software makes

data analysis easy.

1 Plate Setup

2 Protocol Setup

3 Run

96-LED arrayfor illumination

Optics preciselyfocus light onindividual wells

PMTs amplifysignal through

11 stages

Filters allow only thelonger wavelengths

associated withemitted fluorescence

to reach the PMTs 96-well plateatop DNA Engine

thermal cycler

Real-Time Thermal Cycling

© 2004 MJ Research, Inc. 10725 rev A.A

Opticon™ Systems Specifications

Opticon 2 Opticon

Linear Dynamic Range of Starting Copy Number: Up to ten orders of magnitude Up to ten orders of magnitude

Detection Limit of Template Starting Copy Number: Down to single-copy detection Down to single-copy detection

Sample Volume Range: 10–100µl (20µl recommended) 10–100µl (20µl recommended)

Fluorescence Excitation Range: 470–505nm 450–495nm

Fluorescence Detection Range: Ch1: 523–543nm; Ch2: 540–700nm 515–545nm

Input Power: 100-240 Volts AC, 50-60 Hertz, 850W maximum 100-240 Volts AC, 50-60 Hertz, 850W maximum

Size: 34cm wide x 47cm deep x 60cm high 34cm wide x 47cm deep x 60cm high

Weight: 29kg 27kg

DNA Engine® Specifications

Sample Capacity: 96-well microplate (low-profile) or12x0.2ml 8-strip tubes (low-profile)

Thermal Uniformity: ±0.4°C within 30 seconds of arrival at90°C

Thermal Accuracy: Average temp within ±0.3°C of pro-grammed value at 90°C, NIST-traceable

Speed of Ramping: Up to 3°C/second

Thermal Range: 4°C to 105°C

Gradient Performance

Temperature Gradient Accuracy: ±0.3°C of target at end columns within30 seconds of arrival at programmedtarget, NIST-traceable

Thermal Column Uniformity: ±0.4°C, in column, well-to-well, within30 seconds of target attainment

Accuracy of Calculated (displayed) ±0.4°C of actual column temperature, Well Temperature: NIST-traceable

Lowest Temp for Gradient: 30°C

Highest Temp for Gradient: 105°C

Temperature Differential Range: 1.0°C to 24.0°C

Opticon and Opticon Monitor are trademarks, and DNA Engine, DNA Engine Opticon, andhelix logo are trademarks registered in the US, belonging to MJ Research, Incorporated.

SYBR is a registered trademark of Molecular Probes, Inc. FAM is a trademark and VIC is a registered trademark of Applera Corp.

The Polymerase Chain Reaction (PCR) is a process covered by US Patents #4,683,195 and#4,683,202, which are owned by Hoffmann-La Roche. Users should obtain a license to per-form the reaction, and a license is currently available through either Roche Molecular Systemsof Pleasanton, California; or F. Hoffman-La Roche, Ltd. of Basel, Switzerland.

System Specifications

AA homozygote

AGheterozygote

GG homozygote

Automated scoring of genotypes. Differently labeled, SNP-specific hydrolysisprobes were used to detect amplification of different alleles from >200 patient samples.For each sample, the C(t)values obtained with each probe were plotted against eachother (left), using an analysis feature in the Opticon Monitor software. Data plotted in thisway cluster into genotypic groups depending on which channel(s) detected fluorescence.The panels on the right were used to create and define data groups.

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Broad Linear Dynamic Range with Single Copy Detection. Left: Log-fluorescence vs. cycle number for duplicate amplifications of plasmid containing β-actincDNA. Initial template copy number ranged over 10 orders of magnitude. The right plotshows these replicates plotted on a standard curve (automatically generated by OpticonMonitor™ software). The high r2 value (above trace), indicates an excellent linear fit.