Nanopore-based sequencing using voltage mode with hybrid mode stimuli

Abstract

A liquid voltage is applied to a first side of a lipid bilayer. The liquid voltage comprises a tag-reading period with a tag-reading voltage that tends to capture a tag into a nanopore in the lipid bilayer and an open-channel period with an open-channel voltage that tends to repel the tag. A pre-charging voltage source is connected to an integrating capacitor and a working electrode on a second side of the lipid bilayer during a pre-charging time period, such that the integrating capacitor and the working electrode are charged to a pre-charging voltage. The pre-charging voltage source is disconnected from the integrating capacitor and the working electrode during an integrating time period, such that a voltage of the integrating capacitor and a voltage of the working electrode may vary as a current flows through the nanopore. The pre-charging time period overlaps with a beginning portion of the tag-reading period.

1 . A method of analyzing a molecule in a nanopore, comprising:
 applying a liquid voltage to an electrolyte on a first side of a membrane, wherein the liquid voltage comprises a molecule-reading period with a molecule-reading voltage level that that is configured to pull the molecule or a portion of the molecule into a nanopore in the membrane and an open-channel period with an open-channel voltage level that is configured to repel the molecule or the portion of the molecule from the nanopore in the membrane;   connecting a pre-charging voltage source to a capacitor and a working electrode on a second side of the membrane during a pre-charging time period, such that the capacitor and the working electrode are charged to a pre-charging voltage; and   disconnecting the pre-charging voltage source from the capacitor and the working electrode during the molecule-reading period, such that a voltage of the capacitor and a voltage of the working electrode may vary as a current flows through the nanopore in the membrane;   wherein the pre-charging time period spans across a transition from the open channel period to the molecule-reading period.   
     
     
         2 . The method of  claim 1 , wherein the pre-charging time period overlaps with an ending portion of the open-channel period and a beginning portion of the molecule-reading period. 
     
     
         3 . The method of  claim 1 , wherein the pre-charging time period overlaps with a beginning portion of the open-channel period. 
     
     
         4 . The method of  claim 1 , wherein the pre-charging time period overlaps with an ending portion of the molecule-reading period. 
     
     
         5 . The method of  claim 1 , wherein the pre-charging time period spans across a transition from the molecule-reading period to the open-channel period. 
     
     
         6 . The method of  claim 1 , wherein the pre-charging time period has a duration of 100 microseconds to 50 milliseconds. 
     
     
         7 . The method of  claim 1 , wherein the pre-charging time period has a duration of 50 microseconds to 25 milliseconds. 
     
     
         8 . A system, instrument or device for analyzing a molecule, comprising:
 a voltage source;   a counter electrode coupled to the voltage source;   a pre-charging voltage source;   a capacitor that is configurable to connect with the pre-charging voltage source;   a working electrode that is configurable to connect with the pre-charging voltage source; and   a controller configured to:   control the voltage source to apply a liquid voltage via the counter electrode to an electrolyte on a first side of a membrane, wherein the liquid voltage comprises a molecule-reading period with a molecule-reading voltage level that is configured to pull the molecule or a portion of the molecule into a nanopore in the membrane and an open-channel period with an open-channel voltage level that is configured to repel the molecule or the portion of the molecule from the nanopore in the membrane;   connect the pre-charging voltage source to the capacitor and the working electrode on a second side of the membrane during a pre-charging time period, such that the capacitor and the working electrode are charged to a pre-charging voltage;   disconnect the pre-charging voltage source from the capacitor and the working electrode during the molecule-reading period, such that a voltage of the capacitor and a voltage of the working electrode may vary as a current flows through the nanopore in the membrane; and   wherein the pre-charging time period spans across a transition from the open channel period to the molecule-reading period; and   a memory coupled to the controller and configured to provide the controller with instructions.   
     
     
         9 . The system, instrument or device of  claim 8 , wherein the pre-charging time period overlaps with an ending portion of the open-channel period and a beginning portion of the molecule-reading period. 
     
     
         10 . The system, instrument or device of  claim 8 , wherein the pre-charging time period overlaps with a beginning portion of the open-channel period. 
     
     
         11 . The system, instrument or device of  claim 8 , wherein the pre-charging time period overlaps with an ending portion of the molecule-reading period. 
     
     
         12 . The system, instrument or device of  claim 8 , wherein the pre-charging time period spans across a transition from the molecule-reading period to the open-channel period. 
     
     
         13 . The system, instrument or device of  claim 8 , wherein the pre-charging time period has a duration of 100 microseconds to 25 milliseconds.