Single-molecule nanofet sequencing systems and methods

Abstract

Real time electronic sequencing devices, chips, and systems are described. Arrays of nanoFET devices are used to provide sequence information about a template nucleic acid in a polymerase-template complex bound to the nanoFET. The nanoFET devices typically have a source, a drain and a gate comprising a nanowire. A single polymerase enzyme complex comprising a polymerase enzyme complexed with the template nucleic acid is bound to the gate. The polymerase is bound to the gate non-covalently through a polymeric binding agent that has two strands, each strand interacting with the nanowire such that the polymerase is in a central location between the strands with the polymeric binding agent extending away from the polymerase complex along the nanowire in both directions.

1 .- 20 . (canceled) 
     
     
         21 . A nanoFET device comprising:
 a substrate comprising a pair of nanoscale electrodes;   a nanotube extending across the pair of nanoscale electrodes;   a conductive material on top of each of the nanoscale electrodes and on top of the nanotube;   an insulating material over the nanoscale electrodes whereby at least a portion of the nanotube is not covered by the insulating material.   
     
     
         22 . The device of claim  1  wherein the conductive material comprises aluminum, silver, gold, and platinum. 
     
     
         23 . The device of claim  1  wherein the insulating material is a metal oxide or metal nitride. 
     
     
         24 . The device of claim  1  wherein the nanotube comprises a carbon nanotube. 
     
     
         25 . The device of claim  1  wherein the nanotube has a polymerase enzyme bound to it. 
     
     
         26 . The device of claim  1  wherein the length of the nanotube is less than 300 nm. 
     
     
         27 . The device of claim  1  wherein the insulating material comprises a well that constitutes the portion of the nanotube that is not covered by the insulating material. 
     
     
         28 . The device of  claim 27  wherein the well is cylindrical and has a diameter of less than 300 nm. 
     
     
         29 . The device of  claim 27  wherein the well is cylindrical and has a diameter of less than 200 nm. 
     
     
         30 . The device of  claim 27  wherein the well is cylindrical and has a diameter of less than 100 nm. 
     
     
         31 . The device of  claim 27  wherein the ratio of the depth to the width of the well is from 1:2 to 1:10. 
     
     
         32 . The device of  claim 27  wherein the ratio of the depth to the width of the well is from 1.5:1 to 5:1. 
     
     
         33 . The device of claim  7  wherein the depth of the well is between 5 nm and 300 nm. 
     
     
         34 . The device of claim  1  comprising 1000 nanoFETs to a ten million nanoFETs. 
     
     
         35 . The device of claim  1  wherein the substrate comprises silicon. 
     
     
         36 . A chip for sequencing a plurality of single nucleic acid template molecules comprising:
 a plurality of nanoFET devices of  claim 25 :   wherein the substrate is configured such that the plurality of nanoFET devices come into contact with a sequencing reaction mixture comprising a plurality of types of nucleotide analogs each having a different conductivity label; and   a plurality of electrical connection sites for bringing current and voltage to the nanoFETs, and for receiving electrical signals from the nanoFETs.   
     
     
         37 . The chip of  claim 36  wherein the substrate comprises about 1,000 nanoFET devices to about 10 million nanoFET devices. 
     
     
         38 . The chip of  claim 36  wherein the substrate comprises about 10,000 nanoFET devices to about 1 million nanoFET devices. 
     
     
         39 . The chip of  claim 36  wherein at least one of the conductivity labels comprise a polymer chain having multiple charges. 
     
     
         40 . The chip of  claim 36  wherein at least one of the conductivity labels comprise a protein.