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    Method for isolating nucleic acids from a food sample
    22.
    发明授权
    Method for isolating nucleic acids from a food sample 有权

    公开(公告)号：US10011827B2

    公开(公告)日：2018-07-03

    申请号：US14380314

    申请日：2013-02-28

    申请人： QIAGEN GmbH

    发明人： Janina Cramer ,  Sarah Fakih ,  Corinna Küppers ,  Holger Engel

    IPC分类号： C12N15/10 ,  C12N1/06 ,  C12Q1/6806 ,  C12Q1/689

    CPC分类号： C12N15/1003 ,  C12N1/06 ,  C12N15/1013 ,  C12Q1/6806 ,  C12Q1/689

    摘要： The method for isolating nucleic acids from a food sample comprising the following steps: a) obtaining a food enrichment culture; b) transferring a portion of the food enrichment culture into a reaction vessel thereby providing a food enrichment sample and providing a water-immiscible phase in contact with the food enrichment culture; c) lysing the food enrichment sample to provide a lysed sample; d) isolating nucleic acids from the lysed sample. Food enrichment culture samples are known to contain high concentrations of organic and/or liposoluble inhibitors. By contacting the enrichment culture sample with a water-immiscible phase before the actual DNA extraction procedure starts, part of the lipophilic inhibitors is expected to cross the phase interface due to an enhanced solubility in the organic phase and thereby become depleted. The water-immiscible phase provided according to the invention interacts directly with the sample material throughout bacteria lysis and DNA extraction thereby optimizing the subsequent DNA purification processes due to the depletion of food-borne inhibitors. This method yields nucleic acids, in particular DNA that is of an improved quality and purity to be used in a subsequent PCR reaction to detect pathogen nucleic acids in the isolated nucleic acids.

    Method for isolating nucleic acids
    24.
    发明授权
    Method for isolating nucleic acids 有权

    公开(公告)号：US09695465B2

    公开(公告)日：2017-07-04

    申请号：US14238625

    申请日：2012-08-13

    申请人： Thorsten Voss

    发明人： Thorsten Voss

    IPC分类号： G01N1/34 ,  C12Q1/68 ,  C12N15/10

    CPC分类号： C12Q1/6806 ,  C12N15/1003 ,  C12Q2527/125 ,  C12Q2527/127

    摘要： The present invention pertains to a method for isolating nucleic acids from a sample, preferably a blood sample, comprising the following steps: a) obtaining a sample which has been stabilized by the use of at least one cationic detergent, wherein the cationic detergent has formed complexes with the nucleic acids; b) obtaining the complexes optionally together with other sample components from the stabilized sample, wherein said complexes comprise the nucleic acids to be isolated; c) resuspending the complexes and optionally adding one or more additives before, during and/or after resuspension, thereby obtaining a resuspended sample comprising at least: i) the nucleic acid to be isolated; ii) at least one chaotropic agent; and iii) at least one chelating agent; and d) isolating nucleic acids from the resuspended sample. It was found that adding a chelating agent during resuspension considerably increases the nucleic acid yield as the formation of precipitates which irreversibly adhere to the container wall is considerably reduced.

    Method for purifying a target nucleic acid
    25.
    发明授权
    Method for purifying a target nucleic acid 有权

    公开(公告)号：US09624253B2

    公开(公告)日：2017-04-18

    申请号：US13810406

    申请日：2011-07-15

    申请人： Thorsten Voss ,  Ralf Wyrich

    发明人： Thorsten Voss ,  Ralf Wyrich

    IPC分类号： C07H1/08 ,  C12N15/10

    CPC分类号： C07H1/08 ,  C12N15/1003 ,  C12N15/1006

    摘要： The present invention pertains to method for purifying at least a target nucleic acid from a sample, said method comprising at least the following steps: a) incubating the sample with at least one protein-degrading compound; b) binding the target nucleic acid to a solid phase; c) eluting the target nucleic acid from the solid phase; d) incubating the eluted target nucleic acid with at least one protein-degrading compound; e) binding the target nucleic acid again to a solid phase; f) optionally eluting the bound target nucleic acid from the solid phase. It was surprisingly found that performing a second protein digestion step after the target nucleic acid was bound and eluted from a solid phase before the nucleic acids are rebound to a solid phase is very efficient in reducing remaining protein contaminations in the isolated nucleic acid.

    Reaction mixtures for forming cDNA from an RNA template
    27.
    发明授权
    Reaction mixtures for forming cDNA from an RNA template 有权

    公开(公告)号：US09611506B2

    公开(公告)日：2017-04-04

    申请号：US14939153

    申请日：2015-11-12

    申请人： QIAGEN GMBH

    发明人： Holger Engel ,  Subrahmanyam Yerramilli ,  Martin Kreutz ,  Dirk Loeffert ,  Christian Korfhage

    IPC分类号： C12P19/34 ,  C12Q1/68

    CPC分类号： C12Q1/6806 ,  C12Q1/6848 ,  C12Q2527/101 ,  C12Q2521/131 ,  C12Q2521/107

    摘要： This invention relates to a process for synthesis of a cDNA in a sample, in an enzymatic reaction, whereby the process comprises the steps: simultaneous preparation of a first enzyme with polyadenylation activity, a second enzyme with reverse transcriptase activity, a buffer, at least one ribonucleotide, at least one deoxyribonucleotide, an anchor oligonucleotide; addition of a sample that comprises a ribonucleic acid; and incubation of the agents of the previous steps in one or more temperature steps, which are selected such that the first enzyme and the second enzyme show activity. The invention further relates to a reaction mixture that comprises a first enzyme with polyadenylation activity, a second enzyme with reverse transcriptase activity, optionally a buffer, optionally at least one ribonucleotide, optionally at least one deoxyribonucleotide, and optionally an anchor oligonucleotide. Moreover, the invention relates to a kit that comprises a corresponding reaction mixture.

    Method of isolating purified RNA with reduced DNA contaminations
    28.
    发明授权
    Method of isolating purified RNA with reduced DNA contaminations 有权
    用减少的DNA污染物分离纯化的RNA的方法

    公开(公告)号：US09487550B2

    公开(公告)日：2016-11-08

    申请号：US13821159

    申请日：2011-09-06

    申请人： Gabriele Christoffel

    发明人： Gabriele Christoffel

    IPC分类号： C07H21/00 ,  C07H1/08 ,  C12N15/10 ,  C07H19/06 ,  C07H19/16 ,  C07H19/073 ,  C07H19/167 ,  C07H1/06 ,  C07H19/067 ,  C07H19/04

    CPC分类号： C07H1/08 ,  C07H1/06 ,  C07H19/04 ,  C07H19/06 ,  C07H19/067 ,  C07H19/073 ,  C07H19/16 ,  C07H19/167 ,  C12N15/1003

    摘要： The present invention pertains to a method of isolating RNA from a sample comprising RNA, and DNA, comprising: a) adding an acidic denaturing composition comprising a chaotropic agent and phenol to the sample; b) adding a water-insoluble organic solvent and separating the resulting phases thereby forming a multi-phase mixture comprising an aqueous phase, optionally an interphase and an organic phase, wherein the RNA is concentrated in said aqueous phase and DNA and proteins are concentrated in said organic phase and/or in said interphase; and c) isolating said RNA from said aqueous phase, wherein at least one cationic detergent is added before separating the phases. It was found that the addition of at least one cationic detergent considerably reduces the amount of DNA in the aqueous, RNA containing phase. Therefore, the present invention allows to easily isolate pure RNA which comprises considerably less DNA contaminations.

    摘要翻译： 本发明涉及从包含RNA和DNA的样品中分离RNA的方法，其包括：a）向样品中加入包含离液剂和苯酚的酸性变性组合物; b）加入水不溶性有机溶剂并分离所得的相，从而形成包含水相（任选地相间相和有机相）的多相混合物，其中RNA浓缩在所述水相中，并将DNA和蛋白质浓缩 所述有机相和/或所述相间; 和c）从所述水相中分离所述RNA，其中在分离相之前加入至少一种阳离子去污剂。 发现添加至少一种阳离子洗涤剂显着地减少含水RNA的相中的DNA的量。 因此，本发明允许容易地分离包含相当少的DNA污染物的纯RNA。

    Method and device for isolating and purifying double-stranded nucleic acids
    29.
    发明授权
    Method and device for isolating and purifying double-stranded nucleic acids 有权
    用于分离和纯化双链核酸的方法和装置

    公开(公告)号：US09458452B2

    公开(公告)日：2016-10-04

    申请号：US13884416

    申请日：2011-11-08

    申请人： Thorsten Singer ,  Holger Wedler

    发明人： Thorsten Singer ,  Holger Wedler

    IPC分类号： C07H21/00 ,  C12N15/10 ,  B01D15/34 ,  B01D15/38 ,  C07H1/08

    CPC分类号： C12N15/101 ,  B01D15/34 ,  B01D15/3828 ,  C07H1/08

    摘要： The invention relates to a chromatographic device for isolating and/or purifying double-stranded nucleic acids, preferably double-stranded DNA, from a mixture of such nucleic acids with single-stranded nucleic acids, oligonucleotides, mononucleotides, salts and/or other such impurities. The invention also relates to a method for chromatographically isolating and/or purifying same, and to a kit for this purpose.

    摘要翻译： 本发明涉及用于从这种核酸与单链核酸，寡核苷酸，单核苷酸，盐和/或其它这些杂质的混合物中分离和/或纯化双链核酸，优选双链DNA的色谱装置 。 本发明还涉及用于色谱分离和/或纯化的方法以及用于该目的的试剂盒。

    Process for parallel isolation and/or purification of RNA and DNA
    30.
    发明授权
    Process for parallel isolation and/or purification of RNA and DNA 有权
    平行分离和/或纯化RNA和DNA的方法

    公开(公告)号：US09422542B2

    公开(公告)日：2016-08-23

    申请号：US13580474

    申请日：2011-02-25

    申请人： Vera Holländer

    发明人： Vera Holländer

    IPC分类号： C07H21/00 ,  C12N15/10

    CPC分类号： C12N15/1003

    摘要： The present invention relates to a process for the parallel isolation and/or purification of RNA and DNA from the same fixed biological sample, the quantification and analysis of the nucleic acids isolated by the process according to the invention, to a kit for the parallel isolation and/or purification of RNA and DNA from a fixed sample and to the use of this kit for the diagnosis, prognosis, decision with respect to therapy and/or the monitoring of the therapy of a disease.

    摘要翻译： 本发明涉及用于将来自相同固定生物样品的RNA和DNA平行分离和/或纯化的方法，将通过本发明的方法分离的核酸定量和分析用于并行隔离的试剂盒 和/或来自固定样品的RNA和DNA的纯化，以及使用该试剂盒进行治疗和/或对疾病治疗的监测的诊断，预后决定。