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公开(公告)号：US09587263B2
公开(公告)日：2017-03-07
申请号：US14225887
申请日：2014-03-26
Applicant: General Electric Company
Inventor： Ryan Charles Heller , John Richard Nelson , Paresh Lakhubhai Patel , Alison Myfanwy Wakefield , Stephen James Capper
CPC classification number: C12P19/34 , C12Q1/6844 , C12Q2527/107 , C12Q2527/125 , C12Q2527/137 , C12Q2531/119
Abstract: Provided herein are methods and kits for isothermal nucleic acid amplifications that use a target nucleic acid template; a reaction mixture comprising a DNA polymerase having a strand displacement activity, a deoxyribonucleoside triphosphate (dNTP) mixture, a primer with a 3′ end and a 5′ end, a molecular crowding reagent, and a buffer solution for amplifying the target nucleic acid template. The buffer solution maintains a low salt concentration of the reaction mixture, and wherein the salt concentration results in a melting temperature (Tm) of the primer at least 10° C. below the reaction temperature. The amplification is effected under isothermal condition.
Abstract translation: 本文提供了使用靶核酸模板的等温核酸扩增的方法和试剂盒; 包含具有链置换活性的DNA聚合酶，脱氧核糖核苷三磷酸（dNTP）混合物，具有3'端和5'末端的引物，分子拥塞试剂和用于扩增靶核酸模板的缓冲溶液的反应混合物 。 缓冲溶液维持反应混合物的低盐浓度，其中盐浓度导致底漆的熔融温度（Tm）低于反应温度至少10℃。 在等温条件下进行扩增。
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公开(公告)号：US20160053307A1
公开(公告)日：2016-02-25
申请号：US14933275
申请日：2015-11-05
Applicant: GENERAL ELECTRIC COMPANY
Inventor： Ryan Charles Heller , Erik Leeming Kvam , John Richard Nelson
IPC: C12Q1/68
CPC classification number: C12Q1/6844 , C12Q1/6869 , C12Q2521/501 , C12Q2525/307 , C12Q2527/137 , C12Q2531/125
Abstract: Provided herein are methods for generation and amplification of a single-stranded DNA circle in a single reaction vessel from a linear DNA without any intervening purification steps. The single-stranded DNA circle is generated via a template-independent single-stranded DNA ligation. Whole-genome amplification of linear chromosomal DNA in a single tube using ligation-assisted DNA amplification is also provided.
Abstract translation: 本文提供了在单个反应容器中从线性DNA产生和扩增单链DNA环的方法，而没有任何介入的纯化步骤。 通过不依赖模板的单链DNA连接产生单链DNA环。 还提供了使用连接辅助的DNA扩增在单个管中的全基因组扩增线性染色体DNA。
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公开(公告)号：US09217167B2
公开(公告)日：2015-12-22
申请号：US13952040
申请日：2013-07-26
Applicant: General Electric Company
Inventor： Ryan Charles Heller , John Richard Nelson , Erik Leeming Kvam
CPC classification number: C12P19/34 , C12Q1/6869 , C12Q2521/501 , C12Q2525/307 , C12Q2527/137
Abstract: Provided herein are methods for generation and amplification of a single-stranded DNA circle in a single reaction vessel from a linear DNA without any intervening purification steps. The single-stranded DNA circle is generated via a template-independent single-stranded DNA ligation. Whole-genome amplification of circulating nucleic acids extracted from blood is provided. Kits for performing the disclosed methods are also provided.
Abstract translation: 本文提供了在单个反应容器中从线性DNA产生和扩增单链DNA环的方法，而没有任何介入的纯化步骤。 通过不依赖模板的单链DNA连接产生单链DNA环。 提供了从血液中提取的循环核酸的全基因组扩增。 还提供了用于执行所公开的方法的套件。
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公开(公告)号：US10655167B2
公开(公告)日：2020-05-19
申请号：US15949061
申请日：2018-04-09
Applicant: GENERAL ELECTRIC COMPANY
Inventor： Ryan Charles Heller , Erik Leeming Kvam , John Richard Nelson
IPC: C12P19/34 , C12Q1/6844 , C12Q1/6869
Abstract: Provided herein are methods for generation and amplification of a single-stranded DNA circle in a single reaction vessel from a linear DNA without any intervening purification steps. The single-stranded DNA circle is generated via a template-independent single-stranded DNA ligation. Whole-genome amplification of linear chromosomal DNA in a single tube using ligation-assisted DNA amplification is also provided.
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Patent Agency Ranking
公开(公告)号：US10443094B2
公开(公告)日：2019-10-15
申请号：US15419560
申请日：2017-01-30
Applicant: GENERAL ELECTRIC COMPANY
Inventor： Ryan Charles Heller , John Richard Nelson , Paresh Lakhubhai Patel , Alison Myfanwy Wakefield , Stephen James Capper , Peter James Tatnell , Jeffrey Kenneth Horton
IPC: C12Q1/68 , C12Q1/6844 , C12P19/34
Abstract: Provided herein are methods and kits for isothermal nucleic acid amplifications that use a target nucleic acid template; a reaction mixture comprising a DNA polymerase having a strand displacement activity, a deoxyribonucleoside triphosphate (dNTP) mixture, a primer with a 3′ end and a 5′ end, a molecular crowding reagent, and a buffer solution for amplifying the target nucleic acid template. The buffer solution maintains a low salt concentration of the reaction mixture, and wherein the salt concentration results in a melting temperature (Tm) of the primer at least 10° C. below the reaction temperature. The amplification is effected under isothermal condition.
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公开(公告)号：US09938568B2
公开(公告)日：2018-04-10
申请号：US14933275
申请日：2015-11-05
Applicant: GENERAL ELECTRIC COMPANY
Inventor： Ryan Charles Heller , Erik Leeming Kvam , John Richard Nelson
CPC classification number: C12Q1/6844 , C12Q1/6869 , C12Q2521/501 , C12Q2525/307 , C12Q2527/137 , C12Q2531/125
Abstract: Provided herein are methods for generation and amplification of a single-stranded DNA circle in a single reaction vessel from a linear DNA without any intervening purification steps. The single-stranded DNA circle is generated via a template-independent single-stranded DNA ligation. Whole-genome amplification of linear chromosomal DNA in a single tube using ligation-assisted DNA amplification is also provided.
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公开(公告)号：US20170137874A1
公开(公告)日：2017-05-18
申请号：US15419560
申请日：2017-01-30
Applicant: GENERAL ELECTRIC COMPANY
Inventor： Ryan Charles Heller , John Richard Nelson , Paresh Lakhubhai Patel , Alison Myfanwy Wakefield , Stephen James Capper , Peter James Tatnell , Jeffrey Kenneth Horton
IPC: C12Q1/68
CPC classification number: C12Q1/6844 , C12P19/34 , C12Q2527/107 , C12Q2527/125 , C12Q2527/137 , C12Q2531/119
Abstract: Provided herein are methods and kits for isothermal nucleic acid amplifications that use a target nucleic acid template; a reaction mixture comprising a DNA polymerase having a strand displacement activity, a deoxyribonucleoside triphosphate (dNTP) mixture, a primer with a 3′ end and a 5′ end, a molecular crowding reagent, and a buffer solution for amplifying the target nucleic acid template. The buffer solution maintains a low salt concentration of the reaction mixture, and wherein the salt concentration results in a melting temperature (Tm) of the primer at least 10° C. below the reaction temperature. The amplification is effected under isothermal condition.
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公开(公告)号：US20150275282A1
公开(公告)日：2015-10-01
申请号：US14225887
申请日：2014-03-26
Applicant: General Electric Company
Inventor： Ryan Charles Heller , John Richard Nelson , Paresh Lakhubhai Patel , Alison Myfanwy Wakefield , Stephen James Capper
CPC classification number: C12P19/34 , C12Q1/6844 , C12Q2527/107 , C12Q2527/125 , C12Q2527/137 , C12Q2531/119
Abstract: Provided herein are methods and kits for isothermal nucleic acid amplifications that use a target nucleic acid template; a reaction mixture comprising a DNA polymerase having a strand displacement activity, a deoxyribonucleoside triphosphate (dNTP) mixture, a primer with a 3′ end and a 5′ end, a molecular crowding reagent, and a buffer solution for amplifying the target nucleic acid template. The buffer solution maintains a low salt concentration of the reaction mixture, and wherein the salt concentration results in a melting temperature (Tm) of the primer at least 10° C. below the reaction temperature. The amplification is effected under isothermal condition.
Abstract translation: 本文提供了使用靶核酸模板的等温核酸扩增的方法和试剂盒; 包含具有链置换活性的DNA聚合酶，脱氧核糖核苷三磷酸（dNTP）混合物，具有3'端和5'末端的引物，分子拥塞试剂和用于扩增靶核酸模板的缓冲溶液的反应混合物 。 缓冲溶液维持反应混合物的低盐浓度，其中盐浓度导致底漆的熔融温度（Tm）低于反应温度至少10℃。 在等温条件下进行扩增。
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公开(公告)号：US20150079635A1
公开(公告)日：2015-03-19
申请号：US14027947
申请日：2013-09-16
Applicant: General Electric Company
Inventor： Ryan Charles Heller , John Richard Nelson
IPC: C12P19/34
CPC classification number: C12Q1/6853 , C12P19/34 , C12Q1/6865 , C12Q2521/301 , C12Q2523/31 , C12Q2525/179 , C12Q2531/119 , C12Q2531/125 , C12Q2531/143
Abstract: Provided herein are methods and kits for isothermal nucleic acid amplifications that use an oligocation-oligonucleotide conjugate primer for amplifying a target nucleic acid to generate amplicons. Isothermal DNA amplification methods that employ a strand displacing DNA polymerase and polyamine-oligonucleotide conjugate primer are also provided.
Abstract translation: 本文提供了使用寡聚寡核苷酸共轭引物扩增靶核酸以产生扩增子的等温核酸扩增的方法和试剂盒。 还提供了使用链置换DNA聚合酶和多胺 - 寡核苷酸共轭引物的等温DNA扩增方法。
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