Attachment 2 Specifications CFX Duet System Solicitation 1232SA26Q0496.docx

SPECIFICATIONS

Project CFX Duet Real-Time PCR System (3 units), and accompanying software. Brand name or equal.

1.0 Scope of Work / Specifications Description

CFX Duet Real-Time PCR System Quantity: 3 Unit Type: Each Minimum Requirements:

96-well block, 2-color detection plus FRET, must include base unit and cables.

Compatible with CFX Maestro Software.

CFX Maestro Software 2.3 for Windows PC Quantity: 1 Unit Type: Each Minimum Requirements:

1 license, CFX Maestro 2.3, software for real-time PCR plate setup, data collection, statistics, and graphing of results; for Windows PCs.

Contractor Requirements

2.0 Technical /Additional Requirements

CFX Duet Real-Time PCR System Specifications

· System includes an automated lid that opens, closes, and can apply sealing force to reaction wells.

· System does not have lid or drawer that extends beyond the footprint of the system nor requires additional operating clearance

· Thermal gradient enables optimization of multiple temperatures in a single assay. Temperature differences of up to 24°C front-to-back can be created.

· Peltier-driven thermal cycler features a maximum ramping speed of 5°C/sec, with an average ramp rate of 3.3°C/sec.

· Has a thermal block operational range of 0–100°C.

· Sample block temperature accuracy is +/-0.2°C of programmed target at 90°C, with a uniformity of +/-0.3°C well-to-well within 10 seconds of arrival at 90°C.

· Optical system allows excitation and detection of up to two fluorescent dyes in a single reaction well.

· Optics shuttle independently illuminates and detects fluorescence from each well with the same LED per channel without added fiber optics. The system uses three filtered LEDs for illumination and differentially detects emission using three filtered photodiodes (one for each channel). *

· Fixed optical path, directly over each well, eliminates the need to normalize for positional bias.

· Optics shuttle excites fluorescent dyes with absorption spectra in the 450–535 nm range; optimally detects dyes with emission spectra in the 515–580 nm range.

· One channel is dedicated for FRET and Protein Thermal Shift (Protein Melt) experiments — illuminate dyes with absorption spectra in the 450–490 nm range and detect dyes with emission spectra in the 560–580 nm range.

· System reads all 96 wells in all channels within 12 seconds.

· In fast scan mode, the system reads all 96 wells within 3 seconds.

· It has a linear dynamic range of 10 orders of magnitude for detection of input samples.

· Detects one copy of target sequence in human genomic DNA.

· Reaction volumes from 1–50 μL can be used, 10–25 μL recommended.

· Detects ≤10 fmol of fluorescein.

· System can be integrated with an automation system for hands-free operation.

· Will continue to run and complete a run if the software is stopped or interrupted. This prevents run data from being lost in case if there is unintentional interruption of the software. This function is especially valuable for precious or limited samples, whereby all data is not lost for a run that was interrupted.

· Systems shall have low sample volume requirements. Ideally could use as little as 1ul of sample, but up to 10ul is acceptable due to the small size of the insect tissues available. Additionally, we require multiplexing of samples and need to accommodate multiple targets while retaining the low volume of samples needed due to the small size of the insect tissues available.

· Systems shall have flexibility to use multiple chemistries, not- limited to those of the manufacturer.

· Systems shall not require ROX for gene expression assays.

· Systems shall have small footprint to fit into our overcrowded, small lab space, with dimensions not exceeding 14” x 23” x 15” (W x D x H).

· Warranty: Systems must have a minimum of a 1-year warranty from completion of installation and a complete service manual Warranty Requirements for all components

· Systems must have the training needed for use and set up of system for new employees, cost included in cost of item.

· System shall be compatible with CFX Maestro software

CFX Maestro Software specifications:

· Multiplex amplification and melt curve and end-point analyses can be performed on up to five fluorophores in a single reaction well.

· Multiple experiments can be analyzed, and reports generated from a single plate using the Well Groups feature.

· Additional thermal cycler repeat steps can be added to a GOTO loop.

· Run results can be displayed in a customizable configuration so that multiple panes of information may be viewed in a single window.

· Multiple standard curves can be simultaneously viewed when a common fluorophore is used for multiple targets.

· PCR quantification by standard curve features automated calculations of reaction efficiencies with y-intercept.

· Automated allelic discrimination can use end-point fluorescence or quantification cycle (Cq) values.

· Gene expression analysis can be performed by relative quantity (ΔCq) or normalized expression (ΔΔCq).

· If reaction efficiencies are present, these will be automatically utilized in gene expression analysis.

· Multiple reference genes can be assigned in normalized expression (ΔΔCq) analysis.

· Individual reaction efficiencies can be used in normalized expression (ΔΔCq) analysis.

· Reference Gene Selector Tool displays gene stability for selection of ideal reference genes.

· Up to 5,000 Cq values from different data files can be compared for gene expression analysis.

· Multiplex data can be displayed for up to five fluorophores.

· Data graphs and tables can be exported into Microsoft Excel or printed from software menus.

· License for qbasePLUS software permits further gene expression analysis.

· Embedded reports tool can be customized by the user to export run information, data tables, graphs, and analysis parameters in a specified order to PDF or other file formats.

· Exported data type and format can be customized.

· Integrate with any laboratory information management system (LIMS) using built-in LIMS file management.

· Optional CFX Maestro 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.

· The software can be set up to email run files to a specified user after runs are completed.

· Help information is contextual to the active window in the software.

· Software enables data to be grouped and interpreted by both technical replicates and biological groups.

· Image export options include DPI selection up to 600DPI, choice of any image size, and color selection using RGB specifications.

· Image annotation function allows for automatic p-value annotation and addition of arrows, circles, and text directly onto graph images.

· Software performs t-tests and 1- and 2-way ANOVA calculations.

· Multiple plates can be grouped and analyzed together.

· Software integrates with PrimePCR for run setup and results analysis. PrimePCR Controls Analysis tool provides interpretation of controls in a run such as positive control, reverse-transcriptase check, and genomic DNA contamination.

· Software displays data in multiple formats including bar chart, dot plot, box-and-whisker plot, scatter plot, clustergram, and volcano plot.

· Custom data view allows a user to define the specific information to be displayed in a single view, allowing a user to mouse over a well and see that specific result data

3.0 Place of Delivery

USDA, ARS, SEA, Honey Bee Breeding, Genetics, and Physiology Research Laboratory Attn: Michael Simone-Finstrom 1157 Ben Hur Road Baton Rouge, LA 70820

4.0 Delivery Schedule

1157 Ben Hur Road

30 days after award

1157 Ben Hur Road

30 days after award

5.0 Delivery Point of Contact

Michael Simone-Finstrom, Michael.simonefinstrom@usda.gov, (225) 421-9155.