Sequencing by Direct Electrical Measurements of Polymerase Fluctuations

NIH RePORTER · NIH · R01 · $623,671 · view on reporter.nih.gov ↗

Abstract

PROJECT SUMMARY Our goal is to create a low-cost, high speed single-molecule genome sequencer with long reads, requiring no dyes or labels, with direct electronic readout. The technology has the potential for rapid reads (on the order of an hour per genome) using an integrated circuit chip based on simple two terminal devices. If this potential were to be fully realized, then use of genome sequencing in the clinic with near real-time feedback could become a reality. Our data shows that large-amplitude polymerase fluctuations are associated with polymerase activity. Our first goal is to identify signals associated with each nucleotide incorporation with high accuracy. This would enable sequencing by means of cyclic addition of nucleotides, but with the advantage that homopolymer runs of sequence would be counted directly. Our second aim is to identify the individual nucleotides being incorporated based on the details of the signals generated at each incorporation. This would allow a sequencer to run at the free-running speed of the polymerase in the presence of all four nucleotidetriphosphates, so that a wafer of 10,000 devices could produce a genome's worth of reads in an hour. Our third aim is to develop scalable technology for fabrication of prototype solid-state devices.

Key facts

NIH application ID
10165781
Project number
5R01HG011079-02
Recipient
ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
Principal Investigator
STUART LINDSAY
Activity code
R01
Funding institute
NIH
Fiscal year
2021
Award amount
$623,671
Award type
5
Project period
2020-05-15 → 2023-02-28