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1. 发明授权

US08394584B2 Detecting and sorting methylated DNA using a synthetic nanopore 有权
标题翻译：使用合成纳米孔检测和分选甲基化DNA
公开(公告)号：US08394584B2
公开(公告)日：2013-03-12
申请号：US13133300
申请日：2009-12-18
申请人： Gregory Timp , Winston Timp , Andrew Feinberg , Utkur Mirsaidov
发明人： Gregory Timp , Winston Timp , Andrew Feinberg , Utkur Mirsaidov
IPC分类号： C12Q1/68 , C12P19/34 , C12M1/34 , C07H21/02 , C07H19/00
CPC分类号： C12Q1/6827 , C12Q2565/631
摘要： Provided are methods for detecting, characterizing or separating DNA based on methylation. Heterogeneous DNA populations are separated based on DNA methylation by providing a membrane having a nanopore through which an electric field is applied. DNA of interest is introduced, and for a given threshold voltage across the nanopore, only DNA having a methylation parameter of interest may transit the pore, thereby facilitating detection, characterization, or separation of DNA based on methylation. The methods are optionally used to detect a disease state that is associated with DNA methylation including, but not limited to, cancer.
摘要翻译：提供了用于基于甲基化检测，表征或分离DNA的方法。通过提供具有通过其施加电场的纳米孔的膜，基于DNA甲基化分离不均匀DNA群体。引入目标DNA，对于纳米孔上的给定阈值电压，只有具有目标甲基化参数的DNA才能转移孔，从而便于基于甲基化的DNA的检测，表征或分离。任选地，所述方法用于检测与DNA甲基化相关的疾病状态，包括但不限于癌症。

2. 发明授权

US08748091B2 Characterizing stretched polynucleotides in a synthetic nanopassage 有权
标题翻译：表征合成纳米尺度中的拉伸多核苷酸
公开(公告)号：US08748091B2
公开(公告)日：2014-06-10
申请号：US12971240
申请日：2010-12-17
申请人： Gregory Timp , Winston Timp , Utkur Mirsaidov , Aleksei Aksimentiev , Jeffrey Comer
发明人： Gregory Timp , Winston Timp , Utkur Mirsaidov , Aleksei Aksimentiev , Jeffrey Comer
IPC分类号： C12Q1/68 , C12M1/36 , G01N27/00 , C07H21/04 , G01N27/414 , B01L3/00
CPC分类号： C12Q1/6869 , B01L3/5027 , G01N27/4146 , G01N33/48721
摘要： Methods of trapping a deformed portion of a double-stranded polynucleotide in a membrane nanopassage are provided. In an aspect, the membrane has a nanopassage that defines a confine region, wherein the membrane separates a first fluid compartment from a second fluid compartment, and the nanopassage is in fluid communication with the first and second compartments. A polynucleotide is provided to the first fluid compartment and optionally a threshold voltage for the membrane and the polynucleotide is determined. A driving voltage across the membrane that is greater than the threshold voltage is applied to force a portion of the polynucleotide sequence into the nanopassage confine region, and decreased to a holding voltage bias to trap the polynucleotide portion in the nanopassage confine region. In particular, at least one nucleotide base-pair is fixably positioned in the nanopassage confine volume. In further embodiments, any of the trapping methods are used to characterize or sequence double stranded DNA.
摘要翻译：提供了将双链多核苷酸的变形部分捕获在膜纳米尺度中的方法。在一个方面，膜具有界定限制区域的纳滤密度，其中膜将第一流体隔室与第二流体隔室分隔开，并且该纳米尺度与第一和第二隔室流体连通。将多核苷酸提供给第一流体隔室，并且可选地确定膜的阈值电压，并确定多核苷酸。施加大于阈值电压的膜上的驱动电压以迫使一部分多核苷酸序列进入纳米密度限制区域，并降低至保持电压偏压以将多核苷酸部分捕获在纳米尺度限制区域内。特别地，至少一个核苷酸碱基对固定地定位在纳米尺度限制体积中。在另外的实施方案中，任何捕获方法用于表征或序列双链DNA。

3. 发明申请

US20120040343A1 Detecting and Sorting Methylated DNA Using a Synthetic Nanopore 有权
标题翻译：使用合成纳米孔检测和分选甲基化DNA
公开(公告)号：US20120040343A1
公开(公告)日：2012-02-16
申请号：US13133300
申请日：2009-12-18
申请人： Gregory Timp , Winston Timp , Andrew Feinberg , Utkur Mirsaidov
发明人： Gregory Timp , Winston Timp , Andrew Feinberg , Utkur Mirsaidov
IPC分类号： C12Q1/68 , G01N27/447 , B82Y15/00
CPC分类号： C12Q1/6827 , C12Q2565/631
摘要： Provided are methods for detecting, characterizing or separating DNA based on methylation. Heterogeneous DNA populations are separated based on DNA methylation by providing a membrane having a nanopore through which an electric field is applied. DNA of interest is introduced, and for a given threshold voltage across the nanopore, only DNA having a methylation parameter of interest may transit the pore, thereby facilitating detection, characterization, or separation of DNA based on methylation. The methods are optionally used to detect a disease state that is associated with DNA methylation including, but not limited to, cancer.
摘要翻译：提供了用于基于甲基化检测，表征或分离DNA的方法。通过提供具有通过其施加电场的纳米孔的膜，基于DNA甲基化分离不均匀DNA群体。引入目标DNA，对于纳米孔上的给定阈值电压，只有具有目标甲基化参数的DNA才能转移孔，从而便于基于甲基化的DNA的检测，表征或分离。任选地，所述方法用于检测与DNA甲基化相关的疾病状态，包括但不限于癌症。

4. 发明申请

US20110226623A1 Characterizing Stretched Polynucleotides in a Synthetic Nanopassage 有权
标题翻译：表征合成纳米尺度中的拉伸多核苷酸
公开(公告)号：US20110226623A1
公开(公告)日：2011-09-22
申请号：US12971240
申请日：2010-12-17
申请人： Gregory TIMP , Winston TIMP , Utkur MIRSAIDOV , Aleksei AKSIMENTIEV , Jeffrey COMER
发明人： Gregory TIMP , Winston TIMP , Utkur MIRSAIDOV , Aleksei AKSIMENTIEV , Jeffrey COMER
IPC分类号： G01N27/447 , G01N33/50
CPC分类号： C12Q1/6869 , B01L3/5027 , G01N27/4146 , G01N33/48721
摘要： Methods of trapping a deformed portion of a double-stranded polynucleotide in a membrane nanopassage are provided. In an aspect, the membrane has a nanopassage that defines a confine region, wherein the membrane separates a first fluid compartment from a second fluid compartment, and the nanopassage is in fluid communication with the first and second compartments. A polynucleotide is provided to the first fluid compartment and optionally a threshold voltage for the membrane and the polynucleotide is determined. A driving voltage across the membrane that is greater than the threshold voltage is applied to force a portion of the polynucleotide sequence into the nanopassage confine region, and decreased to a holding voltage bias to trap the polynucleotide portion in the nanopassage confine region. In particular, at least one nucleotide base-pair is fixably positioned in the nanopassage confine volume. In further embodiments, any of the trapping methods are used to characterize or sequence double stranded DNA.
摘要翻译：提供了将双链多核苷酸的变形部分捕获在膜纳米尺度中的方法。在一个方面，膜具有界定限制区域的纳滤密度，其中膜将第一流体隔室与第二流体隔室分隔开，并且该纳米尺度与第一和第二隔室流体连通。将多核苷酸提供给第一流体隔室，并且可选地确定膜的阈值电压，并确定多核苷酸。施加大于阈值电压的膜上的驱动电压以迫使一部分多核苷酸序列进入纳米密度限制区域，并降低至保持电压偏压以将多核苷酸部分捕获在纳米尺度限制区域内。特别地，至少一个核苷酸碱基对固定地定位在纳米尺度限制体积中。在另外的实施方案中，任何捕获方法用于表征或序列双链DNA。

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