DNA sequencing via direct electrical detection with free single nucleotide pairing with polymerase
AssigneeSeagate Technology LLC
InventorThomas Young Chang
Life of this patent
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Abstract
A DNA sequencing device having a first conductor electrically insulated from a second conductor, a voltage source and an amplifier electrically connected in series with the first conductor and the second conductor, a DNA polymerase attached to the first conductor and to the second conductor with matching biotinylated tag molecules, and an electric current monitor. A non-discriminating electrical signal is provided by the polymerase during pairing, which signal can be used as a marker to indicate that transcription is occurring between a single-type of free nucleotide and a base nucleotide of a template DNA strand.
Claims11
A method for DNA sequencing comprising:
- providing a direct-electrical-detection-of-polymerase device having a first conductor electrically insulated from a second conductor, a voltage source and an amplifier electrically connected in series with the first conductor and the second conductor, and a DNA polymerase attached to the first conductor and to the second conductor, the conductors, voltage source, and amplifier forming a circuit;
- introducing a template DNA strand to the polymerase for transcription;
- applying a bias voltage to the circuit and measuring a current signal at an output of the amplifier;
- introducing a first type of free nucleotides to the polymerase and monitoring the current signal for an increase;
- responsive to an increase in the current signal, detecting a pairing of a first type of free nucleotide with a leading nucleotide of the template DNA strand, and responsive to no increase in the current signal, determining no pairing of the first type of free nucleotide with the leading nucleotide of the template DNA strand;
- after determining a pairing or no pairing, removing the first type of free nucleotides from the polymerase;
- and
- introducing a second type of free nucleotides different from the first type of free nucleotides to the polymerase and monitoring the current signal for the increase.
A method for DNA sequencing comprising:
- providing a device having a first conductor electrically insulated from a second conductor, a voltage source and an amplifier, electrically connected in series with the first conductor and the second conductor, and a DNA polymerase attached to the first conductor and to the second conductor via matched biotinylated tag molecules;
- the conductors, voltage source, and amplifier forming a circuit;
- introducing a template DNA strand to the polymerase;
- applying a bias voltage to the circuit and measuring a current signal at an output of the amplifier, the current signal having a baseline level;
- introducing a first type of free nucleotides to the polymerase and monitoring the current signal for an increase to a high level, the current signal increase provided by the polymerase during pairing and being non-discriminating on the type of free nucleotide being transcribed to the template DNA strand;
- and
- responsive to the current signal at the high level, determining a pairing of the first type of nucleotide with a leading nucleotide of the template DNA strand, and responsive to the current signal at the baseline level, determining no pairing of the first type of nucleotide with the leading nucleotide of the template DNA strand.
Dependencies are inferred from the claim text.