Abstract: The present disclosure generally relates to the methods and compositions to efficiently analyze polymer characteristics using nanopore-based assays. Specifically disclosed is a method for generating reference signals for polymer analysis in a nanopore system, wherein the nanopore system has a multi-subunit output signal resolution. The method comprises translocating a reference sequence through a nanopore to generate a plurality of reference output signals, wherein each possible multi-subunit sequence that can determine an output signal appears only once in the reference sequence. The output signals are compiled into a reference map for nanopore analysis of an analyte polymer. Also provided are methods and compositions for calibrating the nanopore system for optimized polymer analysis.

Abstract: The present disclosure generally relates to the methods and compositions to efficiently analyze polymer characteristics using nanopore-based assays. Specifically disclosed is a method for generating reference signals for polymer analysis in a nanopore system, wherein the nanopore system has a multi-subunit output signal resolution. The method comprises translocating a reference sequence through a nanopore to generate a plurality of reference output signals, wherein each possible multi-subunit sequence that can determine an output signal appears only once in the reference sequence. The output signals are compiled into a reference map for nanopore analysis of an analyte polymer. Also provided are methods and compositions for calibrating the nanopore system for optimized polymer analysis.

Abstract: Methods for nanopore-based protein analysis are provided. The methods address the characterization of a target protein analyte, which has a dimension greater than an internal diameter of the nanopore tunnel, and which is also physically associated with a polymer. The methods further comprise applying an electrical potential to the nanopore system to cause the polymer to interact with the nanopore tunnel. The ion current through the nanopore is measured to provide a current pattern reflective of the structure of the portion of the polymer interacting with the nanopore tunnel. This is used as a metric for characterizing the associated protein that does not pass through the nanopore.

Abstract: Example devices include a cis well associated with a cis electrode, a trans well associated with a trans electrode, and a field effect transistor (FET) positioned between the cis well and the trans well. Examples of the field effect transistor (FET) include a fluidic system defined therein. The fluidic system includes a first cavity facing the cis well, a second cavity fluidically connected to the trans well, and a through via extending through the field effect transistor from the first cavity. A first nanoscale opening fluidically connects the cis well and the first cavity, the first nanoscale opening having an inner diameter. A second nanoscale opening fluidically connects the through via and the second cavity, the second nanoscale opening having an inner diameter. The second nanoscale opening inner diameter is larger than the first nanoscale opening inner diameter.

Abstract: Example devices include a cis well associated with a cis electrode, a trans well associated with a trans electrode, and a field effect transistor (FET) positioned between the cis well and the trans well. Examples of the field effect transistor (FET) include a fluidic system defined therein. The fluidic system includes a first cavity facing the cis well, a second cavity fluidically connected to the trans well, and a through via extending through the field effect transistor from the first cavity. A first nanoscale opening fluidically connects the cis well and the first cavity, the first nanoscale opening having an inner diameter. A second nanoscale opening fluidically connects the through via and the second cavity, the second nanoscale opening having an inner diameter. The second nanoscale opening inner diameter is larger than the first nanoscale opening inner diameter.

Abstract: Example devices include a cis well associated with a cis electrode, a trans well associated with a trans electrode, and a field effect transistor (FET) positioned between the cis well and the trans well. Examples of the field effect transistor (FET) include a fluidic system defined therein. The fluidic system includes a first cavity facing the cis well, a second cavity fluidically connected to the trans well, and a through via extending through the field effect transistor from the first cavity. A first nanoscale opening fluidically connects the cis well and the first cavity, the first nanoscale opening having an inner diameter. A second nanoscale opening fluidically connects the through via and the second cavity, the second nanoscale opening having an inner diameter. The second nanoscale opening inner diameter is larger than the first nanoscale opening inner diameter.

Abstract: Methods for nanopore-based protein analysis are provided. The methods address the characterization of a target protein analyte, which has a dimension greater than an internal diameter of the nanopore tunnel, and which is also physically associated with a polymer. The methods further comprise applying an electrical potential to the nanopore system to cause the polymer to interact with the nanopore tunnel. The ion current through the nanopore is measured to provide a current pattern reflective of the structure of the portion of the polymer interacting with the nanopore tunnel. This is used as a metric for characterizing the associated protein that does not pass through the nanopore.

Abstract: Example devices include a cis well associated with a cis electrode, a trans well associated with a trans electrode, and a field effect transistor (FET) positioned between the cis well and the trans well. Examples of the field effect transistor (FET) include a fluidic system defined therein. The fluidic system includes a first cavity facing the cis well, a second cavity fluidically connected to the trans well, and a through via extending through the field effect transistor from the first cavity. A first nanoscale opening fluidically connects the cis well and the first cavity, the first nanoscale opening having an inner diameter. A second nanoscale opening fluidically connects the through via and the second cavity, the second nanoscale opening having an inner diameter. The second nanoscale opening inner diameter is larger than the first nanoscale opening inner diameter.

Abstract: Methods for nanopore-based protein analysis are provided. The methods address the characterization of a target protein analyte, which has a dimension greater than an internal diameter of the nanopore tunnel, and which is also physically associated with a polymer. The methods further comprise applying an electrical potential to the nanopore system to cause the polymer to interact with the nanopore tunnel. The ion current through the nanopore is measured to provide a current pattern reflective of the structure of the portion of the polymer interacting with the nanopore tunnel. This is used as a metric for characterizing the associated protein that does not pass through the nanopore.

Abstract: The present disclosure generally relates to the methods and compositions to efficiently analyze polymer characteristics using nanopore-based assays. Specifically disclosed is a method for generating reference signals for polymer analysis in a nanopore system, wherein the nanopore system has a multi-subunit output signal resolution. The method comprises translocating a reference sequence through a nanopore to generate a plurality of reference output signals, wherein each possible multi-subunit sequence that can determine an output signal appears only once in the reference sequence. The output signals are compiled into a reference map for nanopore analysis of an analyte polymer. Also provided are methods and compositions for calibrating the nanopore system for optimized polymer analysis.

Abstract: The present disclosure generally relates to the methods and compositions to efficiently analyze polymer characteristics using nanopore-based assays. Specifically disclosed is a method for generating reference signals for polymer analysis in a nanopore system, wherein the nanopore system has a multi-subunit output signal resolution. The method comprises translocating a reference sequence through a nanopore to generate a plurality of reference output signals, wherein each possible multi-subunit sequence that can determine an output signal appears only once in the reference sequence. The output signals are compiled into a reference map for nanopore analysis of an analyte polymer. Also provided are methods and compositions for calibrating the nanopore system for optimized polymer analysis.

Abstract: Methods for nanopore-based protein analysis are provided. The methods address the characterization of a target protein analyte, which has a dimension greater than an internal diameter of the nanopore tunnel, and which is also physically associated with a polymer. The methods further comprise applying an electrical potential to the nanopore system to cause the polymer to interact with the nanopore tunnel. The ion current through the nanopore is measured to provide a current pattern reflective of the structure of the portion of the polymer interacting with the nanopore tunnel. This is used as a metric for characterizing the associated protein that does not pass through the nanopore.

Abstract: Methods for nanopore-based protein analysis are provided. The methods address the characterization of a target protein analyte, which has a dimension greater than an internal diameter of the nanopore tunnel, and which is also physically associated with a polymer. The methods further comprise applying an electrical potential to the nanopore system to cause the polymer to interact with the nanopore tunnel. The ion current through the nanopore is measured to provide a current pattern reflective of the structure of the portion of the polymer interacting with the nanopore tunnel. This is used as a metric for characterizing the associated protein that does not pass through the nanopore.

Abstract: Methods for nanopore-based protein analysis are provided. The methods address the characterization of a target protein analyte, which has a dimension greater than an internal diameter of the nanopore tunnel, and which is also physically associated with a polymer. The methods further comprise applying an electrical potential to the nanopore system to cause the polymer to interact with the nanopore tunnel. The ion current through the nanopore is measured to provide a current pattern reflective of the structure of the portion of the polymer interacting with the nanopore tunnel. This is used as a metric for characterizing the associated protein that does not pass through the nanopore.

Abstract: Provided herein are methods and systems pertaining to sequencing units of analytes using nanopores. In general, arresting constructs are used to modify an analyte such that the modified analyte pauses in the opening of a nanopore. During such a pause, an ion current level is obtained that corresponds to a unit of the analyte. After altering the modified analyte such that the modified analyte advances through the opening, another arresting construct again pauses the analyte, allowing for a second ion current level to be obtained that represents a second unit of the analyte. This process may be repeated until each unit of the analyte is sequenced. Systems for performing such methods are also disclosed.

Abstract: The present disclosure generally relates to the methods and compositions to efficiently analyze polymer characteristics using nanopore-based assays. Specifically disclosed is a method for generating reference signals for polymer analysis in a nanopore system, wherein the nanopore system has a multi-subunit output signal resolution. The method comprises translocating a reference sequence through a nanopore to generate a plurality of reference output signals, wherein each possible multi-subunit sequence that can determine an output signal appears only once in the reference sequence. The output signals are compiled into a reference map for nanopore analysis of an analyte polymer. Also provided are methods and compositions for calibrating the nanopore system for optimized polymer analysis.

Abstract: Provided herein are artificial membranes of mycolic acids. The membranes may be unsupported or tethered. These membranes are long lived and highly resistant to electroporation, demonstrating their general strength. The mycolic acid membranes are suitable for controlled studies of the mycobacterial outer membrane and can be used in other experiments, such as nanopore analyte translocation experiments.

Abstract: Provided herein are methods and systems pertaining to sequencing units of analytes using nanopores. In general, arresting constructs are used to modify an analyte such that the modified analyte pauses in the opening of a nanopore. During such a pause, an ion current level is obtained that corresponds to a unit of the analyte. After altering the modified analyte such that the modified analyte advances through the opening, another arresting construct again pauses the analyte, allowing for a second ion current level to be obtained that represents a second unit of the analyte. This process may be repeated until each unit of the analyte is sequenced. Systems for performing such methods are also disclosed.

Abstract: Provided herein are artificial membranes of mycolic acids. The membranes may be unsupported or tethered. These membranes are long lived and highly resistant to electroporation, demonstrating their general strength. The mycolic acid membranes are suitable for controlled studies of the mycobacterial outer membrane and can be used in other experiments, such as nanopore analyte translocation experiments.