CFX96™ Real-Time PCR Detection System

Catalog #185-5096
Specifications for Reference Use in Sole-Source Justifications

Bio-Rad Laboratories, Inc. is the original manufacturer and sole supplier of the
CFX96 real-time PCR detection system.
The CFX96 optical reaction module is an interchangeable real-time PCR detection system
used with the C1000™ thermal cycler. The CFX96 system is capable of generating a
thermal gradient across the 96-well reaction block to support rapid optimization of
real-time PCR assays. The CFX96 system is compatible with many fluorescent
technologies for real-time PCR amplification, and enables analysis of up to five
fluorophores in a single reaction well, or one fluorophore when using the SYBR® Green/
FAM channel or FRET channel options for data acquisition. CFX Manager™ software,
which is included with the CFX96 system, supports real-time PCR applications such as
PCR quantitation, melt-curve analysis, allelic discrimination, and gene expression analysis.
Design and performance specifications of the CFX96 real-time PCR detection system:
1. System can run real-time PCR experiments without being attached to a computer.
2. System includes an automated lid that opens, closes, and can apply sealing force to reaction wells.
3. Thermal gradient enables optimization of multiple temperatures in a single assay. Temperature differences of up
to 24°C front-to-back can be created.
4. Peltier-driven thermal cycler features a maximum ramping speed of 5°C/sec, with an average ramp rate of 3.3°C/sec.
5. Has a thermal block operational range of 0–100°C.
6. Sample block temperature accuracy is ±0.2°C of programmed target at 90°C, with a uniformity of ±0.4°C well-to-well
within 10 seconds of arrival at 90°C.
7. Optical system allows excitation and detection of up to five fluorescent dyes in a single reaction well.
8. Optics shuttle independently illuminates and detects fluorescence from each well. The system uses six filtered LEDs for
illumination and differentially detects emission using six filtered photodiodes (one for each channel).
9. Optics shuttle excites fluorescent dyes with absorption spectra in the 450–684 nm range; detects dyes with emission
spectra in the 515–730 nm range.
10. One channel is dedicated for FRET experiments — illuminate dyes with absorption spectra in the 450–490 nm range
and detect dyes with emission spectra in the 560–580 nm range.
11. System reads all 96 wells in all channels within 12 seconds.
12. In fast scan mode the system reads all 96 wells within 3 seconds.
13. Has a linear dynamic range of 10 orders of magnitude for detection of input samples.
14. Detects 1 copy of target sequence in human genomic DNA.
15. Reaction volumes from 1–50 µl can be used,10–25 µl recommended.
16. Detects ≤10 fmol of fluorescein.
CFX Manager software specifications:
1. Multiplex amplification and melt-curve and end-point analyses can be performed on up to five fluorophores in a single
reaction well.
2. Multiple experiments can be analyzed from a single plate using the Well Groups feature.
3. Additional thermal cycler repeat steps can be added to a GOTO loop.
4. PCR quantitation by standard curve features automated calculations of reaction efficiencies.
5. Automated allelic discrimination can use end-point fluorescence or threshold cycle (CT) values.
6. Gene expression analysis can be performed using relative quantity (∆CT) or normalized expression (∆∆CT).
7. Multiple references genes can be assigned in normalized expression (∆∆CT) analysis.
8. Individual reaction efficiencies can be used in normalized expression (∆∆CT) analysis.
9. Up to 5000 CT values from different data files can be compared for gene expression analysis.
10. Multiplex data can be displayed for up to five fluorophores.
11. Data graphs and tables can be exported into Microsoft Excel or printed from software menus.
12. Embedded reports tool can be customized by the user to export run information, data tables, graphs, and analysis
parameters to PDF or other file formats.
13. Optional CFX Manager software, Security Edition provides secure user log-in, detailed audit trail reporting, and electronic
record-keeping tools for compliance with U.S. FDA 21 CFR Part 11 regulations.

Excel and Microsoft are trademarks of Microsoft Corporation. SYBR is a trademark of Invitrogen Corporation.
Purchase of this instrument conveys a limited non-transferable immunity from suit for the purchaser’s own internal research and development and for use in applied fields
other than Human In Vitro Diagnostics under one or more of U.S. Patents Nos. 5,656,493, 5,333,675, 5,475,610 (claims 1, 44, 158, 160–163 and 167 only), and 6,703,236
(claims 1–7 only), or corresponding claims in their non-U.S. counterparts, owned by Applera Corporation. No right is conveyed expressly, by implication or by estoppel
under any other patent claim, such as claims to apparatus, reagents, kits, or methods such as 5' nuclease methods. Further information on purchasing licenses may be
obtained by contacting the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA.
Bio-Rad’s real-time thermal cyclers are licensed real-time thermal cyclers under Applera’s United States Patent No. 6,814,934 B1 for use in research and for all other
fields except the fields of human diagnostics and veterinary diagnostics.
Bio-Rad’s real-time thermal cyclers are covered by one or more of the following U.S. patents or their foreign counterparts owned by Eppendorf AG: U.S. Patent Nos.
6,767,512 and 7,074,367.
Appearances and specifications are subject to change without notice.

Bio-Rad
Laboratories, Inc.

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