1 / 5

Cyclic DNA Sequencing with Enzyme Beads in PicoTiter™ Plate: A Novel Approach

This innovative method utilizes a PicoTiter™ plate to facilitate cyclic DNA sequencing through a series of proprietary enzymatic reactions. By loading DNA and engineered enzyme beads, we enable the production of pyrophosphate (PPi) and ATP, which drive light generation in real-time. The process includes cyclic additions of nucleotides and a specialized diffusion mechanism for efficient nucleotide incorporation. Apyrase enzyme is employed to neutralize unincorporated nucleotides, thereby ensuring accurate signal processing. This technology enhances sequencing speed and accuracy.

hamish
Download Presentation

Cyclic DNA Sequencing with Enzyme Beads in PicoTiter™ Plate: A Novel Approach

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

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

  2. N’ DNA DNA DNA Polymerase (1) + Pyrophosphate (PPi) N dNTP (2) PPi ATP Sulfurylase ATP + SO42- Adenosine Phosphosulfate (APS) Firefly Luciferase Light (3) ATP CO2 + Oxyluciferin + AMP + PPi + D-luciferin + O2 Apyrase (4) dNTP (or ATP) dNMP (or AMP) + 2Pi Pyrophosphate Sequencing Chemistry • Sequencing by synthesis – Cyclic additions of 4 nucleotides: dTTP → dATP → dCTP → dGTP → dTTP … Proprietary

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

  4. Physico-Chemical Processes on PTP A. Nuc Flow Nuc Diffusion B. Nuc incorporation on DNA → PPi production → Diffuses to Enzyme beads → Cyclic ATP & PPi generation reactions start → light generation starts Nuc Flow PPi/ATP Cyclic Reactions 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 C. Apyrase Flow Apyrase diffusion PPi, ATP diffusion Reaction decay 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 D. Wash Buffer Apyrase diffusion PPi, ATP diffusion Reaction ends Convective transport of nucleotides, APS & D-luc Into flow chamber; diffusive transport into wells Proprietary

  5. Simulation: Enzyme Kinetics & Reagent Transport 0.1*[dNTP] Concentration (mM) Reagent Flow [DNA] 107 DNA [PPI] 10 Million copies of DNA in a well Reaction time ~ 30 seconds [ATP] T (sec) Proprietary

More Related