Digital PCR System

Trade Agreement: NAFTA/AIT/Canada FTAs with Peru/Colombia/Panama Tendering Procedures: Generally only one firm has been invited to bid Attachment: None Non-Competitive Procurement Strategy: Government Objectives Representing Best Interests/Value to Govt Comprehensive Land Claim Agreement: No Vendor Name and Address: Bio-Rad Laboratories (Canada) Ltd. 1329 Meyerside Drive Mississauga Ontario Canada L5T1C9 Nature of Requirements: Digital PCR System (ddPRC)

31029-133315/A Allard, Ken Telephone No. - (204) 983-4920 Fax No. - (204) 983-7796

An ACAN is a public notice indicating to the supplier community that a department or agency intends to award a contract for goods, services or construction to a pre-identified supplier, thereby allowing other suppliers to signal their interest in bidding, by submitting a statement of capabilities. If no supplier submits a statement of capabilities that meets the requirements set out in the ACAN, on or before the closing date stated in the ACAN, the contracting officer may then proceed with the award to the pre-identified supplier.

Background: Molecular genetics and genomics research at National Research Council (NRC) - Saskatoon involves a broad suite of activities. Researchers apply genetic approaches using molecular tools, conduct transgenic studies, investigate plant development and physiology, characterise plant and conduct metagenomics studies of microbial populations. A common essential need in these research projects is to quantify nucleic acids with high precision, sensitivity, and specificity at a high throughput level, as related to determining native- and trans-gene copy number, detecting and quantifying genetic variants and target nucleic acids in complex mixtures, and determining gene expression levels. These investigations often involve very limited amounts of nucleic acid samples, as from micro-dissected tissues. The new emerging Digital PCR (dPCR) technology offers very robust system for absolute quantification of nucleic acids and is required to meet the research needs at PBI/NRC-Saskatoon. Any interested supplier must demonstrate by way of a statement of capabilities that its product/equipment/system meets the following requirements:

Criteria for assessment of the Statement of Capabilities - Minimum Essential Performance Specifications:

1. The dPCR system is of a "benchtop" scale must read samples in standard 96-well format plates to enable incorporation into our existing workflows and use standard thermocyclers already present at NRC-Saskatoon. Existing workflows utilise a temperature gradient PCR step to optimise assay conditions and the requested new dPCR system must be able to incorporate a temperature gradient PCR step. The temperature gradient ability must NOT require any additional pipetting or liquid handling steps as these will introduce additional errors into the experiment.

2. The dPCR system must analyze 96 unique samples in less than 5 hours, including the steps of 1) partitioning of sample assays into 12,000 to 20,000 reaction partitions, 2) PCR amplification and 3) reading and scoring reaction partitions. Note: This requirement is necessary in order to analyze multiple samples at once without introducing potential source of error by single tube analyses. Also, these features are needed to meet sample throughput requirements of the multiple users from several NRC research programs as well as potential industrial partners at Saskatoon.

3. This dPCR system should include droplet generator and droplet reader. The droplet reader must incorporate a 96-well auto-sampler so that unattended operation is possible to maximize sample throughput. 4. The dPCR system must produce 12,000 to 20,000 reaction partitions per sample for 96 unique samples in 30 minutes or less.

5. The dPCR system that is compatible with DNA binding chemistries (e.g. Evagreen) as well as hydrolosis probe chemistry using FAM and/or HEX dyes and must detect fluorescence signals corresponding to those derived from VIC-, HEX-, Evagreen-, or FAM-labeled PCR probes.

6. The dPCR system must distinguish between gene copy number variants of 11 vs. 10 and less using two or less sample assay reactions of one reaction well each (equivalent to one chamber or panel of a reaction chip) with an input of nucleic acid template equivalent to 100 ng of plant or microbse' genomic DNA or less so as to minimise amount of nucleic acid sample size required per assay and cost of consumables. This ability must be achieved without the need for a preamplification step which can cause a significant measurement bias by distorting the abundance and relative presence of target nucleic acids.

7. The dPCR system must enable gene expression analysis and quantification of DNA copies with +/- 10% precision to meet the requirements of the multiple users from several research programs. The system must include gene expression module that can analyze and produce accurate gene expression levels in the sample.

8. The dPCR system must detect a specific genetic variant at a level as low as 0.001% abundance in a sample mixture that includes other genetic variants, and enable a linear quantification of this variant over four orders of magnitude of abundance of this variant in a complex mixture of nucleic acids.

9. The dPCR system must NOT require development of standard curves or need an external calibrator to estimate relative abundance of target nucleic acids, thereby minimizing the number of samples to be prepared and analyzed. Rather, the instrument must generate digital data reflecting absolute quantification of target nucleic acid abundance.

10. The digital PCR system must have a reaction partition volume of 1 nL or less.

11. The relative expanded uncertainty of the dPCR system from the data for 20,000 reaction partitions is required to be 5% or less to achieve the data precision required by the research programs.

12. The dPCR system must generate 12,000 to 20,000 reaction partitions from a 20 microlitre sample reaction volume.

13. The dPCR system must NOT require use of specialized liquid handling equipment or robotic liquid handlers that would typically require specialized training and a dedicated technician to operate since the system will be used by numerous researchers from multiple programs.

14. The dPCR system must generate and assay reaction partition in a manner that minimize the dead volume of the partition holder and reader to