Jump to content
[dismiss]
Molecupedia is a parody encyclopedia written by Molecular Reality Corporation. It is not Wikipedia and is not affiliated with the Wikimedia Foundation. About

History of nanopore sequencing

Page protected
From Molecupedia, the encyclopedia of very small things
Development of reading DNA and proteins as they pass through a nanoscale pore
This article is about the history of the technique. For the technique itself, see Nanopore sequencing.

The history of nanopore sequencing runs from a notebook sketch dated June 25, 1989, as the story is usually told, to a pocket-sized sequencer in users' hands in 2014. Its premise was that a single strand of DNA, driven by an electric field through a pore in a membrane, would change the ionic current through the pore in a way that depends on its sequence. The measurement descends from the Coulter counter, whose 1953 patent covered counting particles, blood cells among them, by the change in current they cause in a constriction. Physiologists then learned to record the current of a single ion channel, and in 1996 single strands of DNA and RNA were detected as they crossed an alpha-hemolysin channel. Reading single bases took motor enzymes that ratchet the strand through the pore at a readable speed and pores with a short sensing region, which were first combined in 2012. After the MinION reached early users in 2014 and a field laboratory in Guinea in 2015, the same kinds of pores were used to read peptides and proteins.

Who had the idea first is disputed in print. In 2016 a history of the field by David Deamer, Mark Akeson and Daniel Branton drew a published reply from John Kasianowicz and Sergey Bezrukov, and the two accounts differ on the order and the importance of events. Molecular Reality Corporation, founded in 2018 as demonpore, inc., places its mechanical pores in the same lineage, and its public guide dates "the dream of sequencing by blockade" to 1996.[1]

Precursors

Counting through a hole

Wallace H. Coulter was granted US patent 2,656,508 on October 20, 1953, for counting particles suspended in a fluid by the change in current they cause in a constricted path, with red blood cells as the example.[2] In 1970 DeBlois and Bean extended the method to polystyrene spheres 90 nm across with single submicron pores etched in plastic sheet.[3] The principle is described in the article on the Coulter principle.

Single channels

Erwin Neher and Bert Sakmann recorded the currents of single ion channels in frog muscle in 1976 and shared the 1991 Nobel Prize in Physiology or Medicine "for their discoveries concerning the function of single ion channels in cells".[4][5] A branch of this work at the National Institutes of Health, in a laboratory headed by Adrian Parsegian, used polymers too large to enter a pore to estimate the physical properties of ion channels.[6][7] In 1994 a paper from that laboratory counted polyethylene glycol molecules moving through the channel of the peptide alamethicin, and its opening paragraph describes the Coulter counter.[8] Its first author later published an article titled "Ion channels as molecular Coulter counters to probe metabolite transport".[9]

The pore

Alpha-hemolysin, the toxin of Staphylococcus aureus, can stay open for long periods in a lipid bilayer, which most channels cannot.[10] According to Kasianowicz and Bezrukov, one of them found in 1988, with two colleagues in London, how to keep it open indefinitely, and their experiments in 1990 with polymers of known size showed that the polymers stayed in the pore 500 to 1,000 times longer than expected. Kasianowicz described the result to Deamer at a workshop at NASA Ames in 1991.[6] Hagan Bayley, whose interest lay in how oligomeric pore proteins such as alpha-hemolysin assemble, had meanwhile begun to ask whether such pores could serve as biosensors of molecules and cations. Kasianowicz and Bezrukov write that one of them traveled to Bayley's laboratory at the Worcester Foundation for Experimental Biology, asked whether it could supply purified alpha-hemolysin, and established a collaboration there.[11][6]

The idea

As usually told

In the version told by Deamer, Akeson and Branton, the idea starts in a notebook. In 1989 Deamer "jotted a seemingly implausible idea", on a page dated June 25, 1989: a single strand of DNA drawn through a membrane's nanoscopic pore by electrophoresis, with a current that would change with each base.[11] At about the same time, they write, George Church, whose interest in scaling up sequencing for the Human Genome Project led him to propose that an electronically monitored phage motor embedded in a bilayer might read double-stranded DNA, went on to pursue other methods. Their account then moves to a visit. In late 1993 Deamer went to the National Institute of Standards and Technology to test whether homopolymers of RNA could be electrophoresed through an alpha-hemolysin pore. When a positive bias above about 80 mV was applied, numerous millisecond blockades appeared; no blockades appeared with a negative bias, as expected for a negatively charged polymer.[11]

Deamer and Branton repeated and extended the experiments in February 1994. Longer RNA polymers gave longer blockades, which suggested that the duration was the time the polymer took to cross. Soon afterward a personal communication from the group solving the crystal structure told them that the channel was between about 1.5 nm and 2.5 nm wide, which implied that single-stranded DNA could pass and double-stranded DNA could not. Kasianowicz tested the idea with a mixture of single- and double-stranded DNA designed at Harvard, and the number of molecules that reached the far chamber, counted by quantitative PCR, matched the number of blockades.[11]

Patent

A patent application naming Church, Deamer, Branton, Richard Baldarelli and Kasianowicz was filed on March 17, 1995 and granted as US 5,795,782 on August 18, 1998, under the title "Characterization of individual polymer molecules based on monomer-interface interactions".[12] The government-support statement cites an NIH grant to Church and an NSF grant to Branton. Its background section cites the Human Genome Project as the motivating example. It names alpha-hemolysin and maltoporin as preferred channels, suggests that a polymerase or other DNA-binding protein might ratchet the DNA to prevent backward movement, and proposes a channel whose limiting aperture is much shorter than its full length so that the current would reflect individual monomers. In one embodiment a polymerase is fused to a porin. Deamer, Akeson and Branton call it "the first nanopore sequencing patent".[12][11]

1996

Kasianowicz, Brandin, Branton and Deamer published their paper on November 26, 1996. It showed that an electric field could drive single-stranded RNA and DNA through a channel in a lipid bilayer, and that each molecule appeared as a transient fall in the ionic current whose duration was proportional to its length. The abstract looked ahead: "With further improvements, the method could in principle provide direct, high-speed detection of the sequence of bases in single molecules of DNA or RNA."[10] In 1 M potassium chloride at 120 mV, the count of blockades matched the count of single-stranded molecules that PCR found in the far chamber.[10] Alpha-hemolysin's crystal structure, published the same year, gave the narrowest point of the channel as 1.4 nm, not the 2.6 nm that the DNA paper had used.[13]

Disputed priority

Kasianowicz and Bezrukov answered the 2016 history in a letter that said it omitted "several key ideas, methods, and experimental results" and contained factual errors. They wrote that sequencing DNA with a nanopore was not implausible, that in 1989 one of them had tried to detect DNA with the voltage-dependent anion channel of mitochondria without a conclusive result because of gating, and that Bayley and Oxford Nanopore had abandoned an exonuclease-based method which, as one of the letter's authors showed in a 2012 analysis, could not work unless cleaved nucleotides were prevented from diffusing away from the pore.[6][14] The authors of the history replied that most of their colleagues had called the idea implausible and many grant review panel members had called it impossible, and that they had not known of the unpublished attempts with the voltage-dependent anion channel.[15]

Kasianowicz, Deamer and Branton are named on both the 1995 patent application and the 1996 paper, and both accounts describe the collaboration as beginning with Deamer's approach to Kasianowicz.

Reading bases

In 1999 Akeson, Branton, Kasianowicz, Brandin and Deamer showed that a pore could tell stretches of different bases apart within one molecule. An RNA strand of 70 cytosines followed by 30 adenines produced two current levels in each blockade. The levels reflected the structure of the two stretches: the cytosines form a narrow helix that crosses the pore as it is, and the adenines form a wider helix that must unwind first. Individual bases were not resolved.[16][11] In 2000 Meller and colleagues, including Branton and Golovchenko, found that a pore could distinguish polynucleotides of similar length and composition that differed only in sequence.[17] A single adenine in a strand of cytosines was identified in 2005, and by 2009 all four bases could be told apart in strands that were held in place in the pore.[18][19]

Holding a strand is not reading a molecule that moves. Even the smallest voltage that drives DNA through a pore moves each base past the sensing region in less than about 1 to 10 µs, and identifying a base from picoampere currents needs more than 100 µs. In 2003 the US National Human Genome Research Institute began a program that eventually put more than $50 million into methods for sequencing a human genome for $1,000 or less, and nanopore groups were among those supported.[11] The strategy that worked, which Deamer, Akeson and Branton date to 1998, the year the patent was granted, was to let a processive enzyme control the strand.[11] The Klenow fragment of a DNA polymerase and E. coli exonuclease I slowed translocation by orders of magnitude but did not give base-by-base stepping. Complexes of the polymerase phi29 stayed on the pore more than 10,000 times longer than those of Klenow, and blocking oligomers let one pore process up to 500 strands.[11][20][21]

Another route was to read nucleotides cut from the strand. In 2009 a group at Oxford showed that a protein pore with a covalently attached adapter molecule could identify unlabeled nucleoside monophosphates, the products an exonuclease would release, with an average accuracy of 99.8%.[22]

Solid-state pores

A pore did not have to be a protein. In 2001 a group at Harvard sculpted a pore in a silicon nitride membrane with a feedback-controlled ion beam and registered single DNA molecules in solution through it.[23] In 2003 the group of Cees Dekker shrank pores to single-nanometer size with an electron beam in a transmission electron microscope, and the Harvard group observed double-stranded DNA and its folding in a "solid-state nanopore microscope".[24][25] In 2010 graphene was used as a membrane less than a nanometer thick.[26] Solid-state pores have not yet become commercial sequencers, and Molecular Reality's guide says so plainly: "No solid state pore has done it commercially yet."[1]

Engineered pores and motors

A motor enzyme sitting on the mouth of a pore in a membrane with a DNA strand passing through it, above a staircase of current levels
Strand sequencing with a motor enzyme, schematic. The enzyme feeds the DNA through the sensing constriction one nucleotide at a time, and the current steps from level to level.

Alpha-hemolysin was not an ideal sensor. Its 5 nm stem put about 12 bases into each current level. In the first nanopore sequencing patent a pore with a shorter sensing region had been suggested, and MspA, a porin from Mycobacterium smegmatis with a constriction about 1.2 nm wide and no more than 0.6 nm long, fitted. Wild-type MspA would not take up DNA. In 2008 a group at the University of Washington reported that mutants with the negative charges around the channel replaced by neutral or positive ones detected single strands, and in 2010 the group separated all four bases with larger current differences than alpha-hemolysin gave.[27][28][11] Even MspA senses more than one base at a time. Molecular dynamics simulations suggest that bases spread over about 1.2 nm contribute to each current level, up to five neighbors influence it, and the most likely sequence is therefore inferred, usually by dynamic programming of the Viterbi kind.[11]

In October 2011 a researcher from the University of California, Santa Cruz traveled to Seattle with a phi29 polymerase, blocking oligomers and a synthetic DNA strand, and with a researcher in the Washington group combined the polymerase with an engineered MspA. They soon obtained the first convincing sequential reads of a template strand, a repeating C-A-T trimer, and the two groups reported it in a joint poster at the Advances in Genome Biology and Technology meeting at Marco Island, Florida, in February 2012. Two papers followed in Nature Biotechnology.[11] The Santa Cruz paper ratcheted DNA templates forward and backward through alpha-hemolysin under the control of phi29 polymerase, at median rates of 2.5 to 40 nucleotides a second, with a registry error of 10% to 24.5% at each position in a single pass.[21] The Seattle paper combined the polymerase with mutant MspA and recorded current levels with median durations near 28 ms and differences of up to 40 pA, which matched six known sequences with readable regions 42 to 53 nucleotides long.[29]

MinION

Oxford Nanopore Technologies was founded in 2005, two years after Bayley moved to the University of Oxford, with colleagues who had experience of commercialization. It licensed DNA strand-sequencing patents in 2008 and began an internal effort in 2010.[11] On February 1, 2012 it said it would sell DNA strand sequencing products "directly to customers within 2012".[30] On February 17, 2012, at the meeting at Marco Island, its chief technology officer presented a sequencer the size of a USB memory stick, the MinION, which the company expected to retail at less than $900.[31] The first MinION devices went to users in June 2014 through a large-scale collaborative program, the MinION Access Program.[11]

The MinION described in the 2016 history weighed about 100 g, and its flow cell held 2,048 protein pores, of which the software monitored 512 at a time. In one laboratory at Santa Cruz the average identity of single-molecule reads to a reference sequence rose from 66% in June 2014 to 85% in November 2014, and a consortium of five laboratories that sequenced the same strain of Escherichia coli reported 2D read accuracies of 92% to 93%, a 2D read being one that combines both strands of a hairpin-joined molecule.[11] In April 2015 a MinION-based laboratory was carried to Guinea in standard airline luggage and used to sequence 142 Ebola virus samples, with results less than 24 hours after receiving a positive sample and sequencing that took as little as 15 to 60 minutes.[32] In 2018 the genome of a human cell line was sequenced and assembled from MinION data, 91.2 Gb in total, with ultra-long reads of up to 882 kb.[33] Researchers at the company also showed in 2018 that its pore arrays could sequence RNA directly, without reverse transcription or amplification, and could detect nucleotide analogs in the RNA.[34]

The identity of the pore was a point of dispute. Kasianowicz and Bezrukov asked in 2016 whether the MinION used alpha-hemolysin, an engineered MspA or a variant of either, and the authors of the history replied that the company had kept the information closely held until very recently.[6][15] On May 26, 2016 Oxford Nanopore announced that the new pore of its R9 chemistry was CsgG, licensed from a laboratory at the VIB research institute in Belgium and from University College London, and that the speed would rise from about 70 to about 250 bases a second per pore.[35] CsgG's natural role is to export the fibers of bacterial biofilms.[36]

Proteins

Main articles: Nanopore protein sequencing and Protein sequencing

A protein has to be unfolded before a pore can read it along its length. In 2013 the unfoldase ClpX moved proteins through an alpha-hemolysin pore.[37] In 2020 an aerolysin pore detected all twenty amino acids carried on a short polycationic carrier.[38] In 2021 a helicase pulled a DNA-peptide conjugate through MspA, a single peptide was read 117 times, and the error in identifying a single amino acid change fell below one in a million when the reads were combined.[39] By 2024 ClpX was moving proteins hundreds of amino acids long through CsgG on a commercial sensor array, in two-residue steps. A pore engineered with charges in its lumen has also drawn linearized proteins against the electrophoretic force.[40][41]

Molecular Reality

Kent Kemmish, the company's founder, was on the founding team of Halcyon Molecular from 2009 to 2012, working on DNA sequencing by electron microscopy. His public CV says that in the fall of 2009 he authored a winning NIH/NHGRI Grand Opportunity grant in collaboration with the Church laboratory at Harvard Medical School, which was the formal awardee, with Halcyon as the principal sub-grantee. Church was the last author of a 2013 paper on pulling single DNA molecules out of water and stretching them in air, to which Kemmish contributed.[42][43] Church is an investor in the company and was the first formal member of its scientific advisory board.[44][45] Molecular Reality describes him as a co-inventor of nanopore sequencing, and the patent that Deamer, Akeson and Branton call the first for nanopore sequencing names five inventors, Church among them.[44][12][11]

Molecular Reality's own US patent 10,908,144, filed on May 9, 2018 and granted on February 2, 2021, describes stacks of nanopores in membranes that can slide against one another, so that moving one membrane changes the shape of the passage.[46] In the company's guide the idea sits at the end of a line: "Coulter counted cells, the biophysicists threaded DNA, and we are trying to give the whole family mechanical control."[1]

Timeline

DateEvent
October 20, 1953US patent 2,656,508 for counting particles by resistance change is granted to Wallace Coulter.
1970DeBlois and Bean count 90 nm spheres through single submicron pores.
1976Neher and Sakmann record currents of single ion channels.
June 25, 1989Date on the notebook page of Deamer's sketch of sequencing by pore, as usually told.
Late 1993Deamer visits Kasianowicz at NIST and sees blockades by RNA in alpha-hemolysin.
March 17, 1995Patent application naming Church, Deamer, Branton, Baldarelli and Kasianowicz is filed.
November 26, 1996Single strands of RNA and DNA are detected crossing alpha-hemolysin.
2001A solid-state pore registers single DNA molecules.
2005Oxford Nanopore Technologies is founded.
2008Engineered MspA detects single strands of DNA.
February 2012Polymerase and MspA read a repeating three-base sequence; the MinION is announced.
2013The unfoldase ClpX moves proteins through alpha-hemolysin.
June 2014MinION devices are released through the MinION Access Program.
April 2015A MinION laboratory is used for Ebola surveillance in Guinea.
May 26, 2016Oxford Nanopore announces that its R9 pore is CsgG.
2018A human genome is sequenced and assembled from MinION data.
2021A single peptide is read 117 times at single-amino acid resolution.
2024ClpX moves proteins hundreds of amino acids long through CsgG on a commercial sensor array.

See also

Notes


References

  1. ^ a b c "Keltar's Field Guide to Molecular Reality". Molecular Reality Corporation. August 2026.
  2. ^ W. H. Coulter (October 20, 1953). "US2656508A: Means for counting particles suspended in a fluid". Google Patents. Retrieved October 25, 2026.
  3. ^ DeBlois, R. W.; Bean, C. P. (1970). "Counting and sizing of submicron particles by the resistive pulse technique". Review of Scientific Instruments 41 (7): 909-916. doi:10.1063/1.1684724.
  4. ^ Neher, E.; Sakmann, B. (1976). "Single-channel currents recorded from membrane of denervated frog muscle fibres". Nature 260 (5554): 799-802. doi:10.1038/260799a0.
  5. ^ "The Nobel Prize in Physiology or Medicine 1991". Nobel Prize Outreach. Retrieved October 25, 2026.
  6. ^ a b c d e Kasianowicz, J. J.; Bezrukov, S. M. (2016). "On "Three decades of nanopore sequencing"". Nature Biotechnology 34 (5): 481-482. doi:10.1038/nbt.3570. PMID 27153275.
  7. ^ Zimmerberg, J.; Parsegian, V. A. (1986). "Polymer inaccessible volume changes during opening and closing of a voltage-dependent ionic channel". Nature 323 (6083): 36-39. doi:10.1038/323036a0.
  8. ^ Bezrukov, S. M.; Vodyanoy, I.; Parsegian, V. A. (1994). "Counting polymers moving through a single ion channel". Nature 370 (6487): 279-281. doi:10.1038/370279a0. PMID 7518571.
  9. ^ Bezrukov, S. M. (2000). "Ion channels as molecular Coulter counters to probe metabolite transport". Journal of Membrane Biology 174 (1): 1-13. doi:10.1007/s002320001026. PMID 10741427.
  10. ^ a b c Kasianowicz, J. J.; Brandin, E.; Branton, D.; Deamer, D. W. (1996). "Characterization of individual polynucleotide molecules using a membrane channel". Proceedings of the National Academy of Sciences 93 (24): 13770-13773. doi:10.1073/pnas.93.24.13770.
  11. ^ a b c d e f g h i j k l m n o p Deamer, D.; Akeson, M.; Branton, D. (2016). "Three decades of nanopore sequencing". Nature Biotechnology 34 (5): 518-524. doi:10.1038/nbt.3423. PMID 27153285.
  12. ^ a b c "US5795782A: Characterization of individual polymer molecules based on monomer-interface interactions". Google Patents. August 18, 1998. Retrieved October 25, 2026.
  13. ^ Song, L.; et al. (1996). "Structure of staphylococcal alpha-hemolysin, a heptameric transmembrane pore". Science 274 (5294): 1859-1866. doi:10.1126/science.274.5294.1859. PMID 8943190.
  14. ^ Reiner, J. E.; et al. (2012). "The effects of diffusion on an exonuclease/nanopore-based DNA sequencing engine". Journal of Chemical Physics 137 (21): 214903. doi:10.1063/1.4766363.
  15. ^ a b Deamer, D.; Akeson, M.; Branton, D. (2016). "Author response to Kasianowicz and Bezrukov". Nature Biotechnology 34 (5): 482. doi:10.1038/nbt.3561. PMID 27153276.
  16. ^ Akeson, M.; Branton, D.; Kasianowicz, J. J.; Brandin, E.; Deamer, D. W. (1999). "Microsecond time-scale discrimination among polycytidylic acid, polyadenylic acid, and polyuridylic acid as homopolymers or as segments within single RNA molecules". Biophysical Journal 77 (6): 3227-3233. doi:10.1016/S0006-3495(99)77153-5.
  17. ^ Meller, A.; Nivon, L.; Brandin, E.; Golovchenko, J.; Branton, D. (2000). "Rapid nanopore discrimination between single polynucleotide molecules". Proceedings of the National Academy of Sciences 97 (3): 1079-1084. doi:10.1073/pnas.97.3.1079.
  18. ^ Ashkenasy, N.; et al. (2005). "Recognizing a single base in an individual DNA strand: a step toward DNA sequencing in nanopores". Angewandte Chemie International Edition 44 (9): 1401-1404. doi:10.1002/anie.200462114.
  19. ^ Stoddart, D.; et al. (2009). "Single-nucleotide discrimination in immobilized DNA oligonucleotides with a biological nanopore". Proceedings of the National Academy of Sciences 106 (19): 7702-7707. doi:10.1073/pnas.0901054106.
  20. ^ Lieberman, K. R.; et al. (2010). "Processive replication of single DNA molecules in a nanopore catalyzed by phi29 DNA polymerase". Journal of the American Chemical Society 132 (50): 17961-17972. doi:10.1021/ja1087612.
  21. ^ a b Cherf, G. M.; et al. (2012). "Automated forward and reverse ratcheting of DNA in a nanopore at 5-Å precision". Nature Biotechnology 30 (4): 344-348. doi:10.1038/nbt.2147.
  22. ^ Clarke, J.; et al. (2009). "Continuous base identification for single-molecule nanopore DNA sequencing". Nature Nanotechnology 4 (4): 265-270. doi:10.1038/nnano.2009.12.
  23. ^ Li, J.; Stein, D.; McMullan, C.; Branton, D.; Aziz, M. J.; Golovchenko, J. A. (2001). "Ion-beam sculpting at nanometre length scales". Nature 412 (6843): 166-169. doi:10.1038/35084037. PMID 11449268.
  24. ^ Storm, A. J.; et al. (2003). "Fabrication of solid-state nanopores with single-nanometre precision". Nature Materials 2 (8): 537-540. doi:10.1038/nmat941.
  25. ^ Li, J.; Gershow, M.; Stein, D.; Brandin, E.; Golovchenko, J. A. (2003). "DNA molecules and configurations in a solid-state nanopore microscope". Nature Materials 2 (9): 611-615. doi:10.1038/nmat965. PMID 12942073.
  26. ^ Garaj, S.; et al. (2010). "Graphene as a subnanometre trans-electrode membrane". Nature 467 (7312): 190-193. doi:10.1038/nature09379.
  27. ^ Butler, T. Z.; Pavlenok, M.; Derrington, I. M.; Niederweis, M.; Gundlach, J. H. (2008). "Single-molecule DNA detection with an engineered MspA protein nanopore". Proceedings of the National Academy of Sciences 105 (52): 20647-20652. doi:10.1073/pnas.0807514106.
  28. ^ Derrington, I. M.; et al. (2010). "Nanopore DNA sequencing with MspA". Proceedings of the National Academy of Sciences 107 (37): 16060-16065. doi:10.1073/pnas.1001831107.
  29. ^ Manrao, E. A.; et al. (2012). "Reading DNA at single-nucleotide resolution with a mutant MspA nanopore and phi29 DNA polymerase". Nature Biotechnology 30 (4): 349-353. doi:10.1038/nbt.2171.
  30. ^ "Oxford Nanopore to present DNA "strand sequencing" at AGBT conference; commercialise directly in 2012". Oxford Nanopore Technologies. February 1, 2012. Retrieved October 25, 2026.
  31. ^ "Oxford Nanopore launches mini sequencer". IP Group plc. February 17, 2012. Retrieved October 25, 2026.
  32. ^ Quick, J.; et al. (2016). "Real-time, portable genome sequencing for Ebola surveillance". Nature 530 (7589): 228-232. doi:10.1038/nature16996. PMID 26840485.
  33. ^ Jain, M.; et al. (2018). "Nanopore sequencing and assembly of a human genome with ultra-long reads". Nature Biotechnology 36 (4): 338-345. doi:10.1038/nbt.4060.
  34. ^ Garalde, D. R.; et al. (2018). "Highly parallel direct RNA sequencing on an array of nanopores". Nature Methods 15 (3): 201-206. doi:10.1038/nmeth.4577. PMID 29334379.
  35. ^ "Update: New "R9" nanopore for faster, more accurate sequencing, and new ten minute preparation kit". Oxford Nanopore Technologies. May 26, 2016. Retrieved October 25, 2026.
  36. ^ Goyal, P.; et al. (2014). "Structural and mechanistic insights into the bacterial amyloid secretion channel CsgG". Nature 516 (7530): 250-253. doi:10.1038/nature13768. PMID 25219853.
  37. ^ Nivala, J.; Marks, D. B.; Akeson, M. (2013). "Unfoldase-mediated protein translocation through an alpha-hemolysin nanopore". Nature Biotechnology 31 (3): 247-250. doi:10.1038/nbt.2503. PMID 23376966.
  38. ^ Ouldali, H.; et al. (2020). "Electrical recognition of the twenty proteinogenic amino acids using an aerolysin nanopore". Nature Biotechnology 38 (2): 176-181. doi:10.1038/s41587-019-0345-2. PMID 31844293.
  39. ^ Brinkerhoff, H.; Kang, A. S. W.; Liu, J.; Aksimentiev, A.; Dekker, C. (2021). "Multiple rereads of single proteins at single-amino acid resolution using nanopores". Science 374 (6574): 1509-1513. doi:10.1126/science.abl4381. PMID 34735217.
  40. ^ Motone, K.; et al. (2024). "Multi-pass, single-molecule nanopore reading of long protein strands". Nature 633 (8030): 662-669. doi:10.1038/s41586-024-07935-7. PMID 39261738.
  41. ^ Sauciuc, A.; et al. (2024). "Translocation of linearized full-length proteins through an engineered nanopore under opposing electrophoretic force". Nature Biotechnology 42 (8): 1275-1281. doi:10.1038/s41587-023-01954-x. PMID 37723268.
  42. ^ "Stanley Kent Kemmish, Jr." Molecular Reality Corporation. May 2026.
  43. ^ "Molecular Threading: Mechanical Extraction, Stretching and Placement of DNA Molecules from a Liquid-Air Interface". PLoS ONE 8 (7): e69058. 2013. doi:10.1371/journal.pone.0069058.
  44. ^ a b "Company records". Molecular Reality Corporation. October 2026.
  45. ^ Kemmishtree (March 29, 2026). "(no subject)". EQBio-OS.
  46. ^ "US10908144B2: Stacks of coaligned nanopores and fabrication and uses thereof". Google Patents. February 2, 2021. Retrieved October 25, 2026.

Further reading

  • Branton, D.; et al. (2008). "The potential and challenges of nanopore sequencing". Nature Biotechnology 26 (10): 1146-1153. doi:10.1038/nbt.1495.
  • Kasianowicz, J. J.; et al. (2008). "Nanoscopic porous sensors". Annual Review of Analytical Chemistry 1: 737-766. doi:10.1146/annurev.anchem.1.031207.112818.
Retrieved from "https://molecularreality.com/wiki/History_of_nanopore_sequencing"
Last edited 7 hours ago by PicoampPedant