A06_Specifications_40605055.pdf

Real-time PCR Specifications

The Whitney Genetics Lab (WGL) requests to purchase a Bio-Rad real-time PCR detection system.

The use of real-time polymerase chain reaction (PCR) assays for the detection and identification of aquatic invasive species, such as in the invasive carp monitoring program, as well as the identification of rare or endangered species is an integral part of the WGL eDNA program. When the lab first began researching and analyzing eDNA the Bio-Rad CFX system was chosen for its ease of use, accuracy, and available software to perform analytical research. Since then, the lab has purchased multiple Bio-Rad CFX Touch Real- Time PCR detection systems. All our markers and reagent research and validations have been performed on the CFX system platform. The reliability and reproducibility of results has been optimized for this BIO-Rad system and is part of the requirement listed in the Quality Assurance Project Plan (QAPP) for the eDNA monitoring program. Bio-Rad has discontinued the CFX Touch Real-Time PCR system and will stop providing support for the discontinued system within the next 4 years so a suitable replacement must be obtained.

The Bio-Rad CFX Opus Real-Time PCR System is designed to replace the Bio-Rad CFX Real- Time System, this new system uses the same CFX platform as the discontinued model. All of the labs reagents and markers and collected data will be compatible and transferable to the new updated version of the CXF Real-Time PCR units.

System Specifications:

• Sample Capacity: 384 wells

• Sample Size: 1-30 µL

• Touch-screen interface: yes

• Communications: USB 2.0

Optical Detection System Specifications:

• Excitation: 5 filtered LEDs

• Detection: 5 filtered photodiodes

• Range of Excitation/Emission Wavelengths: 450-650 nm

• Sensitivity: Detects one (1) copy of target sequence in human genomic DNA

• Multiplex Analysis: Up to five (5) targets per well

Reaction Block Specifications:

• Type: Fixed, 384-well

• Heating and Cooling Method: Peltier

• Lid Heating: 30-110 °C

• Temperature Range: 4-100 °C

• Maximum Ramp Rate: 2.5 °C /sec

• Average Ramp Rate: 2 °C /sec

• Thermal Accuracy °C: ±0.2 of programmed target at 90°C

• Thermal Uniformity °C: ±0.3 well-to-well after 10 sec of arrival at target

• Gradient Operational Range: 30-100 °C

• Programmable Span: 1-24 °C

Software Specifications:

• Operating System: Windows

• Capable of: Plate setup, data collection, statistical analysis, graphing results

Special Requirements

• Manufacturer Requirement: Bio-Rad 384 CFX platform

• Required trade in discount for existing USFWS equipment:

o (2) Bio-Rad CFX 384 Touch Deep Well Real-Time PCR Detection Systems

Delivery Address and Requested Delivery Date:

Whitney Genetics Lab 555 Lester Ave.

Onalaska, WI 54650 Delivery: 03/01/2023

Technical Point of Contact:

ATTN: TBA AT AWARD

U.S. Fish and Wildlife Service Whitney Genetics Lab

PHONE: TBA AT AWARD

EMAIL: TBA AT AWARD