公开(公告)号：US10920312B2

公开(公告)日：2021-02-16

申请号：US14841120

申请日：2015-08-31

申请人： Roche Sequencing Solutions, Inc.

发明人： Morgan Mager ,  John Foster

IPC分类号： C23C14/35 ,  C12Q1/6874 ,  C23C14/06 ,  C23C14/34 ,  G01N33/487

摘要： A nanopore cell is disclosed. The nanopore cell includes an electrolyte well having a bottom base, a surrounding sidewall, and a hydrophobic surface above the surrounding sidewall. The nanopore cell further includes a first layer of electrode material disposed on the bottom base of the electrolyte well. The nanopore cell further includes a second layer of electrode material disposed on the surrounding sidewall of the electrolyte well and electrically connected to the first layer of electrode material. The first layer of electrode material and the second layer of electrode material are configured to jointly provide capacitive coupling when an electrolyte is placed in the electrolyte well.

公开(公告)号：US11740227B2

公开(公告)日：2023-08-29

申请号：US17804445

申请日：2022-05-27

申请人： Roche Sequencing Solutions, Inc.

发明人： John Foster ,  Morgan Mager

IPC分类号： G01N33/487 ,  H01L29/40 ,  G01N27/447

CPC分类号： G01N33/48721 ,  G01N27/44791 ,  H01L29/401

摘要： A nanopore cell includes a conductive layer and a working electrode disposed above the conductive layer and at the bottom of a well into which an electrolyte may be contained, such that at least a portion of a top base surface area of the working electrode is exposed to the electrolyte. The nanopore cell further includes a first insulating wall disposed above the working electrode and surrounding a lower section of a well, and a second insulating wall disposed above the first insulating wall and surrounding an upper section of the well, forming an overhang above the lower section of the well. The upper section of the well includes an opening that a membrane may span across, and wherein a base surface area of the opening is smaller than the at least a portion of the top base surface area of the working electrode that is exposed to the electrolyte.

公开(公告)号：US20220291193A1

公开(公告)日：2022-09-15

申请号：US17804445

申请日：2022-05-27

申请人： Roche Sequencing Solutions, Inc.

发明人： John Foster ,  Morgan Mager

IPC分类号： G01N33/487 ,  H01L29/40 ,  G01N27/447

摘要： A nanopore cell includes a conductive layer and a working electrode disposed above the conductive layer and at the bottom of a well into which an electrolyte may be contained, such that at least a portion of a top base surface area of the working electrode is exposed to the electrolyte. The nanopore cell further includes a first insulating wall disposed above the working electrode and surrounding a lower section of a well, and a second insulating wall disposed above the first insulating wall and surrounding an upper section of the well, forming an overhang above the lower section of the well. The upper section of the well includes an opening that a membrane may span across, and wherein a base surface area of the opening is smaller than the at least a portion of the top base surface area of the working electrode that is exposed to the electrolyte.

公开(公告)号：US10317392B2

公开(公告)日：2019-06-11

申请号：US15632190

申请日：2017-06-23

申请人： Roche Sequencing Solutions, Inc.

发明人： Morgan Mager ,  John Mannion

IPC分类号： G01N33/487 ,  C12Q1/6869 ,  C12Q1/6874 ,  G01N27/447 ,  G01N27/327 ,  G06F19/22 ,  B82Y5/00 ,  B82Y15/00 ,  G01N27/416

摘要： Improved multi-cell nanopore-based sequencing chips and methods can employ formation, characterization, calibration, and/or normalization techniques. For example, various methods may include one or more steps of performing physical checks of cell circuitry, forming and characterizing a lipid layer on the cells, performing a zero point calibration of the cells, forming and characterizing nanopores on the lipid layers of each cell, performing a sequencing operation to accumulate sequencing signals from the cells, normalizing those sequencing signals, and determining bases based on the normalized sequencing signals.

公开(公告)号：US11346836B2

公开(公告)日：2022-05-31

申请号：US16948401

申请日：2020-09-16

申请人： Roche Sequencing Solutions, Inc.

发明人： John Foster ,  Morgan Mager

IPC分类号： H01L29/40 ,  G01N27/447 ,  G01N33/487

摘要： A nanopore cell includes a conductive layer and a working electrode disposed above the conductive layer and at the bottom of a well into which an electrolyte may be contained, such that at least a portion of a top base surface area of the working electrode is exposed to the electrolyte. The nanopore cell further includes a first insulating wall disposed above the working electrode and surrounding a lower section of a well, and a second insulating wall disposed above the first insulating wall and surrounding an upper section of the well, forming an overhang above the lower section of the well. The upper section of the well includes an opening that a membrane may span across, and wherein a base surface area of the opening is smaller than the at least a portion of the top base surface area of the working electrode that is exposed to the electrolyte.

公开(公告)号：US11293062B2

公开(公告)日：2022-04-05

申请号：US16843528

申请日：2020-04-08

申请人： Roche Sequencing Solutions, Inc.

发明人： John Mannion ,  Morgan Mager

IPC分类号： C12Q1/6869 ,  G01N27/327 ,  G01N27/447 ,  G16B40/00 ,  G16B45/00 ,  G16B40/20

摘要： Techniques for measuring sequences of nucleic acids are provided. Time-based measurements (e.g., forming a histogram) particular to a given sequencing cell can be used to generate a tailored model. The model can include probability functions, each corresponding to different states (e.g., different states of a nanopore). Such probability functions can be fit to a histogram of measurements obtained for that cell. The probability functions can be updated over a sequencing run of the nucleic acid so that drifts in physical properties of the sequencing cell can be compensated. A hidden Markov model can use such probability functions as emission probabilities for determining the most likely nucleotide states over time. For sequencing cells involving a polymerase, a 2-state classification between bound and unbound states of the polymerase can be performed. The bound regions can be further analyzed by a second classifier to distinguish between states corresponding to different bound nucleotides.

公开(公告)号：US10809243B2

公开(公告)日：2020-10-20

申请号：US14841127

申请日：2015-08-31

申请人： Roche Sequencing Solutions, Inc.

发明人： John Foster ,  Morgan Mager

IPC分类号： G01N33/487 ,  G01N27/447 ,  H01L29/40

摘要： A nanopore cell includes a conductive layer and a working electrode disposed above the conductive layer and at the bottom of a well into which an electrolyte may be contained, such that at least a portion of a top base surface area of the working electrode is exposed to the electrolyte. The nanopore cell further includes a first insulating wall disposed above the working electrode and surrounding a lower section of a well, and a second insulating wall disposed above the first insulating wall and surrounding an upper section of the well, forming an overhang above the lower section of the well. The upper section of the well includes an opening that a membrane may span across, and wherein a base surface area of the opening is smaller than the at least a portion of the top base surface area of the working electrode that is exposed to the electrolyte.

公开(公告)号：US10648027B2

公开(公告)日：2020-05-12

申请号：US15669207

申请日：2017-08-04

申请人： Roche Sequencing Solutions, Inc.

发明人： John Mannion ,  Morgan Mager

IPC分类号： C12Q1/6869 ,  G16B40/00 ,  G16B45/00 ,  G01N27/447

摘要： Techniques for measuring sequences of nucleic acids are provided. Time-based measurements (e.g., forming a histogram) particular to a given sequencing cell can be used to generate a tailored model. The model can include probability functions, each corresponding to different states (e.g., different states of a nanopore). Such probability functions can be fit to a histogram of measurements obtained for that cell. The probability functions can be updated over a sequencing run of the nucleic acid so that drifts in physical properties of the sequencing cell can be compensated. A hidden Markov model can use such probability functions as emission probabilities for determining the most likely nucleotide states over time. For sequencing cells involving a polymerase, a 2-state classification between bound and unbound states of the polymerase can be performed. The bound regions can be further analyzed by a second classifier to distinguish between states corresponding to different bound nucleotides.

公开(公告)号：US11892444B2

公开(公告)日：2024-02-06

申请号：US17076508

申请日：2020-10-21

申请人： Roche Sequencing Solutions, Inc.

发明人： Morgan Mager ,  John Mannion

IPC分类号： C12Q1/6869 ,  G01N33/487 ,  C12Q1/6874 ,  G01N27/447 ,  G16B20/00 ,  G01N27/327 ,  G16B30/00 ,  B82Y5/00 ,  B82Y15/00 ,  G01N27/416

CPC分类号： G01N33/48721 ,  C12Q1/6869 ,  C12Q1/6874 ,  G01N27/3278 ,  G01N27/44791 ,  G16B20/00 ,  G16B30/00 ,  B82Y5/00 ,  B82Y15/00 ,  C12Q2563/116 ,  C12Q2565/631 ,  G01N27/4163 ,  C12Q1/6869 ,  C12Q2537/165 ,  C12Q2565/631

摘要： Improved multi-cell nanopore-based sequencing chips and methods can employ formation, characterization, calibration, and/or normalization techniques. For example, various methods may include one or more steps of performing physical checks of cell circuitry, forming and characterizing a lipid layer on the cells, performing a zero point calibration of the cells, forming and characterizing nanopores on the lipid layers of each cell, performing a sequencing operation to accumulate sequencing signals from the cells, normalizing those sequencing signals, and determining bases based on the normalized sequencing signals.

公开(公告)号：US11788132B2

公开(公告)日：2023-10-17

申请号：US17683890

申请日：2022-03-01

申请人： Roche Sequencing Solutions, Inc.

发明人： John Mannion ,  Morgan Mager

IPC分类号： C12Q1/6869 ,  G16B30/00 ,  G01N27/447 ,  G16B40/00 ,  G16B45/00 ,  G16B40/20 ,  G01N33/487

CPC分类号： C12Q1/6869 ,  G01N27/44791 ,  G01N33/48721 ,  G16B40/00 ,  G16B40/20 ,  G16B45/00 ,  C12Q1/6869 ,  C12Q2535/122 ,  C12Q2537/165 ,  C12Q2565/631

摘要： Techniques for measuring sequences of nucleic acids are provided. Time-based measurements (e.g., forming a histogram) particular to a given sequencing cell can be used to generate a tailored model. The model can include probability functions, each corresponding to different states (e.g., different states of a nanopore). Such probability functions can be fit to a histogram of measurements obtained for that cell. The probability functions can be updated over a sequencing run of the nucleic acid so that drifts in physical properties of the sequencing cell can be compensated. A hidden Markov model can use such probability functions as emission probabilities for determining the most likely nucleotide states over time. For sequencing cells involving a polymerase, a 2-state classification between bound and unbound states of the polymerase can be performed. The bound regions can be further analyzed by a second classifier to distinguish between states corresponding to different bound nucleotides.