Systems and methods for electronic detection with nanoFETs
There is disclosed a system for electrical charge detection comprising a nanoFET device. Also disclosed is a method of electrical charge detection for single molecule sequencing. The method includes attaching a macromolecule or assemblies thereof to a gate of a nanoFET device and flowing in a solution of charge tags, where a charge tag includes a nucleotide attached to a charge complex. The method also includes incorporating one charge tag into the macromolecule or assemblies thereof and cleaving the charge tags from the macromolecule or assemblies thereof. The method further includes detecting at least one of current and voltage from the nanoFET device.
1. A method of sequencing a nucleic acid strand, the method comprising:
attaching the nucleic acid strand to a surface of a device, the device having at least one nanoFET having a source, a drain, and a gate, the nucleic acid strand disposed over the gate;
applying a primer to hybridize with the nucleic acid strand following attaching the nucleic acid strand to the surface;
following applying the primer, applying a charge labeled probe to ligate to an end of the primer in response to a sequence of the nucleic acid strand, the charge labeled probe including an oligonucleotide probe complementary to the sequence and a charge tag linked to the oligonucleotide probe;
cleaving the charge tag from the ligated charge labeled probe; and
detecting a change in at least one of current or voltage in response to cleaving the charge label.
2. The method of claim 1 , wherein attaching the nucleic acid strand includes coupling the nucleic acid strand to a primer attached to the surface of the device.
3. The method of claim 1 , wherein attaching the nucleic acid strand includes coupling the nucleic acid strand to biotin, the biotin interacting with an avidin coupled to the surface of the device.
4. The method of claim 1 , wherein attaching the nucleic acid strand includes coupling the nucleic acid strand to an enzyme coupled to the surface of the device.
5. The method of claim 4 , wherein the enzyme is a polymerase.
6. The method of claim 1 , wherein the strand of nucleic acids is a strand of target DNA.
7. The method of claim 1 , wherein applying a charge labeled probe includes flowing a solution of charge labeled probes, the method further comprising removing the solution of charge labeled probes and any unligated charge labeled probes prior to cleaving.
8. The method of claim 1 , wherein the primer and the oligonucleotide probe ligate to form a second nucleic acid strand, the method further comprising:
removing the second nucleic acid strand; and
adding a second primer configured to detect bases shifted by one base relative to the primer,
hybridizing said second primer to the nucleic acid strand; and
adding a second solution of charged labeled probes.
9. The method of claim 8 , further comprising:
ligating a second charge labeled probe of the second solution of charge labeled probes;
cleaving a second charge tag from the second charge labeled probe; and
detecting a change in at least one of current or voltage using the nanoFET device in response to cleaving the second charge tag.
10. The method of claim 1 , further comprising applying a voltage bias to drive the charge labeled probe toward or away from the nanoFET device.
11. The method of claim 1 , further comprising applying a voltage bias to drive the charge labeled probes away from the nanoFET device and pull the nucleic acid strand.
12. The method of claim 1 , wherein the charge tag linked to the oligonucleotide probe is photocleavable.
13. The method of claim 12 , wherein cleaving the charge tag includes activating a light source.
14. The method of claim 1 , wherein the charge tag linked to the oligonucleotide probe is chemically cleavable.