SON_MMTD_dPCR.doc_revised 10SEP2020 RFQ.pdf

STATEMENT OF NEED FOR SUPPLY OR SERVICE

PURPOSE:

The Southwest Fisheries Science Center (SWFSC) Marine Mammal and Turtle Division (MMTD) seeks to purchase a two new pieces of genetics laboratory equipment. The first item is a Digital Polymerase Chain Reaction (dPCR) machine. This Digital PCR System, uses a unique water-emulsion technology that fractionates a DNA sample into tens of thousands of droplets.

This method reduces the likelihood of laboratory error by providing an absolute measure of target DNA molecules with accuracy and sensitivity. Additionally, this instrument generates tens of thousands of data points from a single sample as opposed to a single result generated by traditional quantitative PCR machines. The second item is a standard touchscreen Thermal Cycler for PCR capable of operating a temperature gradient across the block. This instrument will allow us to generate data consistent with data generated by the dPCR machine and will simultaneously increase our data production. Obtaining genetic samples from protected species is a challenge. As such, each sample that is collected is extremely rare, valuable, and irreplaceable.

Together these two PCR systems will provide a multifold increase in the data generated from each sample while maintaining consistent and comparable datasets thereby enhancing MMTD’s genomics capabilities. Additionally, this machine will be utilized by the SWFSC’s Fisheries Resources Division (FRD) and will offer similar benefits to their program.

TECHNICAL SPECIFICATIONS:

Thermal Cycler (General Specs):

• Sample capacity: 96 x 0.2ml tubes or strips

• Maximum ramp rate, °C/sec: 4

• Average ramp rate, °C/sec: 2.5

• Temperature range: 4-100°C

• Temperature accuracy: ±0.5°C of programmed target

• Temperature uniformity: ±0.5°C well-to-well within 30 sec of arrival at target temperature

• Input power: 100–150 VAC, 50–60 Hz; 220–240 VAC, 50–60 Hz; 700 W maximum

• Display: 5.7'' VGA color touch screen

• Port: 1 USB A

• Memory: 500 typical programs; unlimited with USB flash drive expansion

• Dimensions (W x D x H): 26 x 47 x 23 cm (10 x 18 x 9'')

• Weight : 9 kg (20 lb)

• Gradient range : 30–100°C

• Temperature differential range: 1–25°C

Digital PCR (dPCR) System- includes the following components:

• Droplet generator, droplet reader, computer, software, associated component consumables, for EvaGreen or probe-based PCR applications.

• PCR Plate Sealer (includes heat sealing instrument, plate support block that holds 96-well and 384-well plates, sealing frame, power cord.

• Thermal Cycler with 96–Deep Well Reaction Module- Modular thermal cycler platform, includes Thermal Cycler chassis, 96-deep well reaction module, USB flash drive

• dPCR Droplet Reader Oil- 2 L (2 x 1 L), oil for use with droplet reader

• Droplet Generation Oil for Probes - 70 ml (10 x 7 ml), oil for use with droplet generator

• dPCR 96-Well Plates - Pkg of 25, clear well/clear shell semi-skirted plates, for use with

• PCR Plate Heat Seal, foil, pierceable Pkg of 100, foil seals for PCR

Additional Specifications:

• Generates partitions a 20ul RCR reaction into ~20,000 nanoliter-sized droplets

• Can accurately count target copies when multiple target copies are distributed in a droplet

• Each droplet is approximately 1nl and has diameter of approximately 120um

• Droplets are generated using microfluidic technologies

• Droplets are PCR amplified in bulk on thermal cycler with 96-well format

• Droplet reader analyzes a full 96-well plate in less than 2.5hrs

• Droplet reader performs excitation and detection of up to two fluorescent dyes/droplet.

FAM, HEX, (or VIC) and EvaGreen Dyes can used

• Droplet Reader illuminates and detects fluorescence from each individual droplet. A system uses two light-emitting diodes for illumination and differentially detects emission using two filtered multi-pixel photon counters (one for each channel).

• Automatically detects fluorescence with both channels during the reading process.

• Droplet Reader detects droplets without the need for additional passive dyes.

• Droplet Reader can automatically gate out poor-quality droplets using algorithms

• dPCR System has a linear dynamic range of 5 orders of magnitude for detection of input samples

• dPCR System can detect one copy of target sequence from DNA samples

• dPCR System enables the distinction between four and five copies of input target nucleic acid between samples

• Precision ±10% can be achieved

• One 96-well plate can be used to analyze approximately 1,500,000 droplets

• dPCR System is compatible with TaqMan probe chemistry

• dPCR System that is compatible with EvaGreen DNA binding chemistry

• dPCR System that is compatible with temperature gradient on a thermal cycler for optimization of the thermal cycling protocol.

• dPCR System does not require an external gas tank or gas compressor

• DNA from unread droplets can be recovered for down-stream applications

1Yr Full On-Site Extended Cover Service Plan

• Unlimited repair

• Inclusive of travel, parts, and labor

• Includes Droplet Generator

• Includes Droplet Reader

• 1 Preventative Maintenance visit included

DELIVERABLE SCHEDULE:

Delivery date by September 30, 2020

TECHNICAL POINT OF CONTACT (TOC): TO BE PROVIDED AT TIME OF AWARD