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公开(公告)号：US20220145383A1
公开(公告)日：2022-05-12
申请号：US17481374
申请日：2021-09-22
发明人： James White , Ruth Moysey , Mihaela Misca
IPC分类号： C12Q1/6869
摘要： The invention relates to a method of characterising a target polynucleotide using a single-stranded binding protein (SSB). The SSB is either an SSB comprising a carboxy-terminal (C-terminal) region which does not have a net negative charge or a modified SSB comprising one or more modifications in its C-terminal region which decreases the net negative charge of the C-terminal region.
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公开(公告)号：US11155860B2
公开(公告)日：2021-10-26
申请号：US14415459
申请日：2013-07-18
发明人： James White , Ruth Moysey , Mihaela Misca
IPC分类号： C12Q1/6869
摘要： The invention relates to a method of characterising a target polynucleotide using a single-stranded binding protein (SSB). The SSB is either an SSB comprising a carboxy-terminal (C-terminal) region which does not have a net negative charge or a modified SSB comprising one or more modifications in its C-terminal region which decreases the net negative charge of the C-terminal region.
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公开(公告)号：US20210180124A1
公开(公告)日：2021-06-17
申请号：US17094571
申请日：2020-11-10
IPC分类号： C12Q1/6869 , G01N27/447 , G01N33/487 , B82Y15/00
摘要： The invention relates to a new method of determining the presence, absence or characteristics of an analyte. The analyte is coupled to a membrane. The invention also relates to nucleic acid sequencing.
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公开(公告)号：US20200024654A1
公开(公告)日：2020-01-23
申请号：US16338399
申请日：2017-09-29
发明人： Andrew John Heron , James Edward Graham , Richard Alexander Gutierrez , Rebecca Victoria Bowen , James White , Clive Gavin Brown , Daniel George Fordham
IPC分类号： C12Q1/6869 , C12Q1/6876 , G01N33/487
摘要： The invention provides a method of detecting a target polynucleotide in a sample comprising: (a) contacting the sample with a guide polynucleotide that binds to a sequence in the target polynucleotide and a polynucleotide-guided effector protein, wherein the guide polynucleotide and polynucleotide-guided effector protein form a complex with any target polynucleotide present in the sample; (b) contacting the sample with a membrane comprising a transmembrane pore; (c) applying a potential to the membrane; and (d) monitoring for the presence or absence of an effect resulting from the interaction of the complex with the transmembrane pore to determine the presence or absence of the complex, thereby detecting the target polynucleotide in the sample.
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公开(公告)号：US20190352709A1
公开(公告)日：2019-11-21
申请号：US16465987
申请日：2017-11-29
发明人： James Clarke , James White , Richard Muscat , Jessica Mary May Knott , Ramiz Iqbal Nathani , Andrew John Heron , Mark John Bruce , Lakmal Nishantha Jayasinghe , Domenico Caprotti , David Jackson Stoddart , Rebecca Victoria Bowen , Christopher James Wright , Paul Richard Moody
IPC分类号： C12Q1/6869 , G01N33/487
摘要： Methods of characterizing an analyte using a nanopore. One aspect features methods for characterizing a double-stranded polynucleotide using a nanopore, e.g., without using a hairpin connecting a template and a complement of the double-stranded polynucleotide. Another aspect features methods for characterizing an analyte using a tag-modified nanopore with increased sensitivity and/or higher throughput. Compositions and systems including, e.g., adaptors for attachment to double-stranded polynucleotides and tag-modified nanopores, which can be used in the methods are also provided.
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公开(公告)号：US09885078B2
公开(公告)日：2018-02-06
申请号：US14455394
申请日：2014-08-08
发明人： Lakmal Jayasinghe , John Hagan Pryce Bayley , Stephen Cheley , Brian McKeown , James White , James Anthony Clarke
CPC分类号： C12Q1/6869 , C07K14/31 , C12N9/1247 , C12N9/1252 , C12N9/127 , C12N9/1276 , C12N9/16 , C12N9/22 , C12N9/52 , C12N9/90 , C12N9/96 , C12Q2565/631
摘要： The invention relates to constructs comprising a transmembrane protein pore subunit and a nucleic acid handling enzyme. The pore subunit is covalently attached to the enzyme such that both the subunit and enzyme retain their activity. The constructs can be used to generate transmembrane protein pores having a nucleic acid handling enzyme attached thereto. Such pores are particularly useful for sequencing nucleic acids. The enzyme handles the nucleic acid in such a way that the pore can detect its component nucleotides by stochastic sensing.
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公开(公告)号：US20160257942A1
公开(公告)日：2016-09-08
申请号：US15028651
申请日：2014-09-10
发明人： Mark Bruce , Andrew John Heron , Ruth Moysey , Szabolcs Soeroes , Elizabeth Jayne Wallace , James White
摘要： The invention relates to a new method of characterising a target polynucleotide. The method uses a pore and a Dda helicase. The helicase controls the movement of the target polynucleotide through the pore. The invention also relates to modified Dda helicases which can be used to control the movement of polynucleotides and are particularly useful for sequencing polynucleotides.
摘要翻译： 本发明涉及表征靶多核苷酸的新方法。 该方法使用孔和Dda解旋酶。 解旋酶控制靶多核苷酸通过孔的运动。 本发明还涉及可用于控制多核苷酸运动的修饰的Dda解旋酶，并且特别可用于测序多核苷酸。
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公开(公告)号：US09551023B2
公开(公告)日：2017-01-24
申请号：US14427554
申请日：2013-09-06
发明人： Daniel John Turner , Clive Gavin Brown , Stuart William Reid , James Anthony Clarke , James White , David Jackson Stoddart
CPC分类号： C12Q1/6806 , C07H21/04 , C12Q1/6869 , C12Q2525/119 , C12Q2525/301 , C12Q2531/125 , C12Q2533/107 , C12Q2565/631
摘要： The invention relates to a method for modifying a template polynucleotide for characterization, especially for nanopore sequencing. The method produces a modified polynucleotide which is complementary to the template polynucleotide at some positions and which contains universal or abasic nucleotides at the other, and in some instances predicable, positions. The resulting modified polynucleotide can then be characterized.
摘要翻译： 本发明涉及用于修饰模板多核苷酸进行表征的方法，特别是用于纳米孔测序。 该方法产生修饰的多核苷酸，其在某些位置与模板多核苷酸互补，并且在另一个位置包含通用或非碱基核苷酸，并且在某些情况下可以是可预测的位置。 然后可以表征所得修饰的多核苷酸。
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公开(公告)号：US20210018500A1
公开(公告)日：2021-01-21
申请号：US16916305
申请日：2020-06-30
发明人： Daniel John Turner , Daniel George Fordham , Roger Charles Gill , Clive Gavin Brown , Stuart Reid , James Anthony Clarke , James White
IPC分类号： G01N33/543 , G01N33/53 , C12N15/115
摘要： The invention relates to a new method of determining in a sample the presence or absence of one or more analyte members of a group of two or more analytes. The invention therefore relates to a multiplex assay for determining the presence or absence of each analyte in a group of multiple analytes. The assay uses aptamers and transmembrane pores.
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公开(公告)号：US20190249231A1
公开(公告)日：2019-08-15
申请号：US16343580
申请日：2017-10-20
IPC分类号： C12Q1/6825 , C12Q1/6876
CPC分类号： C12Q1/6825 , C12Q1/68 , C12Q1/6816 , C12Q2523/307 , C12Q2525/101 , C12Q2525/161 , C12Q2537/143 , C12Q2563/116 , C12Q2565/607 , C12Q2565/631
摘要： A method for determining the presence, absence or amount of two or more target polynucleotides in a sample comprising additional components, the method comprising: (i) contacting the sample with a panel of two or more probes under conditions suitable for hybridisation of the target polynucleotides to the probes, wherein: (a) each probe comprises a non-hybridisation region and a hybridisation region that specifically hybridises to one of the target polynucleotides to form a hybridised probe; and (b) the hybridisation region of a probe of the panel comprises one or more non-natural nucleotides; (ii) contacting the sample prepared in step (i) with a transmembrane pore through which a single stranded polynucleotide but not a double stranded polynucleotide can pass and applying a potential difference to the transmembrane pore such that the hybridised probes in the sample interact with the pore; (iii) measuring current blockades having a duration within a defined window, wherein: (a) the one or more non-natural nucleotides present in the hybridisation region of the probe increase or decrease the duration of the current blockade due to the probe hybridised to its target polynucleotide such that the proportion of current blockades that occur within the window due to the interaction of the hybridised probes with the pore is increased compared to when the corresponding one or more natural nucleotides are present in the hybridisation region; and (b) each hybridised probe gives rise to a current blockade indicative of that probe; and (iv) correlating the measured current blockades with the probes, thereby determining the presence, absence or amount of the two or more target polynucleotides in the sample.
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