In the project Definition Phase, all mutually agreed settings for the project are documented into an assay design input document. Arrangements from the contract and customer-required specifications serve as input information. In addition, the risk assessment plan and (live) documentation are now initiated.
Finally, an overall assay project plan is compiled for set-up of:

• the assay design strategy, the plan for information and genetic analyses, in order to select the most promising targets for the Design Phase.
• the plan for the Development Phase (till design freeze).
• the plan for the Verification Phase.

The Design Phase is used to prepare the assay design strategy document, built around the settings agreed to in the Definition Phase and assay project plan. This in silico design phase makes use of external resources (e.g. literature, GenBank) and internal know-how (e.g. design software) to select the raw materials for starting the Development Phase.

The actual wet lab assay development starts here. Based on the Design Phase, all steps for the assay build-up are carefully performed and reported. To exemplify, for a qPCR multiplex assay the routine build-up is from monoplex to multiplex for the primers and subsequently for the probes. Each primer/probe selection is followed by optimization of concentration and finalized by determination of ratios there-of. Output of the Development Phase is a design freeze.

At the start of this Verification Phase, first a prototype batch will be prepared, according to the design freeze formulation. Using this prototype batch, the complete requirement specification (design input) will be addressed and tested. These customer-agreed specifications are leading, but next to that, NYtor always starts internally with a routine QC1 (in-kit) / QC2 (key samples) check.

During the Transfer Phase all relevant output documents are combined in the Assay Design History file package.
Subsequent steps are as follows:

• the assay design history file serves as input for the final project review, to be performed together with the customer.
• following a successful final project review, the design and corresponding documentation is transferred (possibly also including a set of prototype kits).
• a close-out project review is carried out to monitor the key performance indicators (e.g. time) and customer satisfaction/complaints, all in support of management reviews and improvement cycles.