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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: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:US10597691B2
公开(公告)日:2020-03-24
申请号:US15408509
申请日:2017-01-18
Applicant: General Electric Company
Inventor: Ryan Charles Heller , John Richard Nelson , Paresh Lakhubhai Patel , Alison Myfanwy Wakefield , Stephen James Capper
IPC: C12P19/34 , C12Q1/6844
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:US20170121747A1
公开(公告)日:2017-05-04
申请号:US15408509
申请日:2017-01-18
Applicant: General Electric Company
Inventor: Ryan Charles Heller , John Richard Nelson , Paresh Lakhubhai Patel , Alison Myfanwy Wakefield , Stephen James Capper
IPC: C12P19/34
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.
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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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