1. Proprietary Bead Deposition Load DNA & Enzyme beads into PicoTiter™ Plate

2. Proprietary Pyrophosphate Sequencing Chemistry  Sequencing by synthesis – Cyclic additions of 4 nucleotides: dTTP → dATP → dCTP → dGTP → dTTP … DNA Polymerase dNTP DNA N Pyrophosphate (PPi) DNA + (1) Adenosine Phosphosulfate (APS) ATP Sulfurylase PPi+ SO 4 2- ATP(2) dNTP (or ATP) Apyrase dNMP (or AMP) + 2Pi(4) N’ ATP CO 2 + Oxyluciferin + AMP + PPi + Firefly Luciferase D-luciferin + O 2 (3) Light

3. Proprietary Simultaneous Enzymatic Reactions in Hundreds of Thousands of Picoliter-size Wells

4. Proprietary Physico-Chemical Processes on PTP A. Nuc Flow Nuc Diffusion Convective transport of nucleotides, APS & D-luc Into flow chamber; diffusive transport into wells B. Nuc Flow PPi/ATP Cyclic Reactions Nuc incorporation on DNA → PPi production → Diffuses to Enzyme beads → Cyclic ATP & PPi generation reactions start → light generation starts C. Apyrase Flow Reaction decay Apyrase diffusion PPi, ATP diffusion PPi, ATP start diffusing out of well; Apyrase flows into chamber & diffuses into well → Neutralizes unincorporated nucs & ATP → Cyclic reaction starts to decay → signal starts to decay D. Wash Buffer Reaction ends Apyrase diffusion PPi, ATP diffusion Wash Buffer flows into chamber; Unused apyrase diffuses out of well → PPi, ATP continue to diffuse out of well → Reaction comes to an end → signal decays to baseline

5. Proprietary T (sec) Concentration (  M) 0.1*[dNTP] [PPI] [ATP] [DNA] 10 7 DNA Reagent Flow 10 Million copies of DNA in a well Reaction time ~ 30 seconds Simulation: Enzyme Kinetics & Reagent Transport