1. Specify gene and state precisely what will be tested (foundation of assay development) 2. Identify the source of the gene (what type of samples will be used to extract DNA for the assay). This factors in quantity, accessibility and concentration to develop workflow 3. Determine the stability of the gene during the course of the assay so assay development is not compromised 4. Determine number of samples included in assay. The more samples the more need for automated systems 5. Determine how the results will be measured. Will it be: qualitative (pass/fail or positive/negative), Semi-quantitative (more graduations than a pass or fail and indicate results on a general scale) or Quantitative measurements (correct and exact numeric measurement)