公开(公告)号：US20160160198A1

公开(公告)日：2016-06-09

申请号：US15048624

申请日：2016-02-19

Applicant: General Electric Company

Inventor： John Richard Nelson ,  Robert Scott Duthie ,  Gregory Andrew Grossman ,  Anuradha Sekher

IPC: C12N9/22 ,  C12Q1/68

CPC classification number: C12N9/22 ,  C07K1/00 ,  C07K14/00 ,  C12Q1/6806 ,  C12Q1/6844 ,  C12Q1/6858 ,  C12Y301/26 ,  C12Q2521/301 ,  C12Q2525/101 ,  C12Q2537/1376 ,  C12Q2527/101

Abstract: Provided herein are mutant endonuclease V enzymes that are capable of nicking an inosine-containing DNA sequence. Nucleic acid assays and agents that employ such mutant endonuclease V enzymes to introduce a nick into a target DNA including one or more inosine, and uses a DNA polymerase to generate amplicons of a target DNA are also described.

Abstract translation: 本文提供了能够切割含有肌苷的DNA序列的突变体内切核酸酶V酶。 还描述了使用这种突变体内切核酸酶V酶将切口引入到包括一种或多种肌苷的靶DNA中并使用DNA聚合酶产生靶DNA扩增子的核酸测定法和试剂。

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

IPC: C12Q1/68 ,  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.

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。

公开(公告)号：US20150167065A1

公开(公告)日：2015-06-18

申请号：US14106264

申请日：2013-12-13

Applicant: General Electric Company

Inventor： John Richard Nelson ,  David Roger Moore ,  Bing Li ,  Robert Scott Duthie ,  Patrick McCoy Spooner

IPC: C12Q1/68

CPC classification number: C12Q1/6844 ,  C12Q1/6853 ,  C12Q2521/307 ,  C12Q2531/119

Abstract: Provided herein are methods for amplification a target dsDNA that is impregnated within a porous matrix using endonuclease-assisted DNA amplification. The amplicons may be subsequent detected within the porous matrix or may be eluted out of the porous matrix. Methods for extracting a genetic material from a biological sample using endonuclease-assisted DNA amplification within a porous matrix are also provided.

Abstract translation: 本文提供了使用内切核酸酶辅助的DNA扩增来扩增浸渍在多孔基质内的靶dsDNA的方法。 可以在多孔基质内随后检测扩增子，或者可以从多孔基质中洗脱。 还提供了在多孔基质内使用内切核酸酶辅助的DNA扩增从生物样品中提取遗传物质的方法。

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。

公开(公告)号：US20150004611A1

公开(公告)日：2015-01-01

申请号：US13929976

申请日：2013-06-28

Applicant: General Electric Company

Inventor： Anthony John Murray ,  John Richard Nelson

IPC: C12P19/34

CPC classification number: C12P19/34 ,  C12N15/111 ,  C12N15/115 ,  C12N2310/16 ,  C12N2320/10

Abstract: Methods for developing a binding-element are provided. A mixture comprising a target molecule, a plurality of oligonucleotides and a ligase is provided, followed by binding the oligonucleotides to the target molecule to form an oligonucleotides-target molecule complex. The oligonucleotides bound to the target molecule are ligated to form the binding-element. The binding-elements are separated from the mixture.

Abstract translation: 提供了开发结合元件的方法。 提供包含靶分子，多个寡核苷酸和连接酶的混合物，然后将寡核苷酸与靶分子结合以形成寡核苷酸 - 靶分子复合物。 与靶分子结合的寡核苷酸被连接以形成结合元件。 结合元件与混合物分离。

公开(公告)号：US20130130352A1

公开(公告)日：2013-05-23

申请号：US13729566

申请日：2012-12-28

Applicant: GENERAL ELECTRIC COMPANY

Inventor： John Richard Nelson ,  Robert Scott Duthie ,  Gregory Andrew Grossman

IPC: C12N9/12

CPC classification number: C12N9/1241 ,  C12Q1/6848 ,  C12Q2525/125 ,  C12Q2521/319

Abstract: Methods and kits for generating contamination-free reagents and reagent solutions for use in nucleic acid amplification are provided. Methods include processing of polymerase solutions, nucleotide solutions and primer solutions to render contaminating nucleic acid inert. The methods employ the proofreading activity of the polymerase and/or exonucleases to de-contaminate the reagents and reagent solutions. Methods and kits for contamination-free nucleic acid amplification are provided.