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    • 1. 发明申请
    • DETECTION OF MULTIPLE NUCLEIC ACID SEQUENCES IN A REACTION CARTRIDGE
    • 在反应盒中检测多个核酸序列
    • WO2010128041A1
    • 2010-11-11
    • PCT/EP2010/056030
    • 2010-05-04
    • QIAGEN GMBHROTHMANN, ThomasENGEL, HolgerHIMMELREICH, RalfWENDE, AndyDAHLKE, Rainer
    • ROTHMANN, ThomasENGEL, HolgerHIMMELREICH, RalfWENDE, AndyDAHLKE, Rainer
    • C12Q1/68
    • C12Q1/6818Y02A50/59C12Q2527/107C12Q2537/143C12Q2565/629
    • The present invention relates to a method for amplifying and detecting nucleic acid sequences in a reaction cartridge comprising the following steps, (i) providing a sample comprising at least one nucleic acid molecule, (ii) in a first reaction chamber of the cartridge providing reagents for an amplification reaction, (iii) mixing the sample with the amplification reagents, (iv) amplifying the at least one nucleic acid in the first reaction chamber of the cartridge, (v) transferring at least parts of the amplification reaction into a second and third reaction chamber of the cartridge each comprising a probe set, wherein (a) each probe set consists of at least three probes, (b) each of the probes is specific for a nucleic acid sequence, (c) there are at least two probes in each set which carry an identical label, (d) each of the probes in a given probe set that carries an identical label has a melting temperature (Tm) which differs by more than 2°C from the other probe in said probe set with the same label, (e) wherein the probes carrying the identicai label differ in melting temperature (Tm) in a way that they are distinguishable by melting point, (f) performing a melting point analysis in order to determine which of the probes has specifically bound a nucleic acid.
    • 本发明涉及用于扩增和检测反应盒中的核酸序列的方法,包括以下步骤:(i)提供包含至少一个核酸分子的样品,(ii)在所述盒的第一反应室中提供试剂 用于扩增反应,(iii)将样品与扩增试剂混合,(iv)扩增筒的第一反应室中的至少一种核酸,(v)将至少部分扩增反应转移到第二和 每个探针组由至少三个探针组成,(b)每个探针对于核酸序列是特异性的,(c)存在至少两个探针 在每个携带相同标签的组中,(d)给定探针组中携带相同标记物的每个探针的熔解温度(Tm)与所述探针中的另一个探针相差大于2℃ (e)其中携带同一标签的探针的熔解温度(Tm)不同,其熔点可区分,(f)进行熔点分析以确定哪个探针 已经特异地结合核酸。
    • 3. 发明申请
    • SIMULTANEOUS DETECTION OF MULTIPLE NUCLEIC ACID SEQUENCES IN A REACTION
    • 在反应中同时检测多个核酸序列
    • WO2009135832A1
    • 2009-11-12
    • PCT/EP2009/055404
    • 2009-05-05
    • Qiagen GmbHQiagen Hamburg GmbHROHTHMANN, ThomasENGEL, Holger
    • ROHTHMANN, ThomasENGEL, Holger
    • C12Q1/68
    • C12Q1/6818C12Q2527/107C12Q2537/143
    • The present invention relates to a method for simultaneously amplifying and detecting nucleic acid sequences in a reaction comprising the following steps: (i) providing a sample comprising at least one nucleic acid molecule; (ii) providing reagents for performing an amplification reaction, wherein the reagents comprise at least four, preferably at least five, more preferably at least six probes, wherein (a) each of the probes is specific for a nucleic acid sequence; (b) at least two, preferably at least three probes carry the same label; and (c) each of the probes that carry the same label has a melting temperature (Tm) which differs by more than 2°C from the other probes with the same label when they are dissociated from their target nucleic acid sequence by heating; (iii) amplifying the nucleic acid sequences in the reaction; (iv) detecting the amplified nucleic acids by determining whether the labeled probe has bound its nucleic acid sequence; and (v) detecting the temperature at which each given labeled probe dissociates from the nucleic acid sequence to which it has bound. The invention also relates to kits for the use in such a method.
    • 本发明涉及在反应中同时扩增和检测核酸序列的方法,包括以下步骤:(i)提供包含至少一个核酸分子的样品; (ii)提供用于进行扩增反应的试剂,其中试剂包含至少四个,优选至少五个,更优选至少六个探针,其中(a)每个探针对核酸序列是特异性的; (b)至少两个,优选至少三个探头携带相同的标签; 和(c)携带相同标记物的每个探针当通过加热从其靶核酸序列解离时,具有与具有相同标记的其它探针相差大于2℃的解链温度(Tm); (iii)扩增反应中的核酸序列; (iv)通过确定标记的探针是否结合其核酸序列来检测扩增的核酸; 和(v)检测每个给定的标记探针从其所结合的核酸序列解离的温度。 本发明还涉及用于这种方法的试剂盒。
    • 5. 发明申请
    • VERFAHREN ZUR SYNTHESE EINER CDNA IN EINER PROBE IN EINER ENZYMATISCHEN REAKTION
    • METHOD FOR A cDNA在一个示例合成在酶反应
    • WO2009021757A1
    • 2009-02-19
    • PCT/EP2008/051763
    • 2008-02-13
    • QIAGEN GMBHENGEL, HolgerYERRAMILLI, SubrahmanyamKREUTZ, MartinLÖFFERT, DirkKORFHAGE, Christian
    • ENGEL, HolgerYERRAMILLI, SubrahmanyamKREUTZ, MartinLÖFFERT, DirkKORFHAGE, Christian
    • C12Q1/68
    • C12Q1/6806C12N15/1096C12Q1/6844C12Q2527/101C12Q2521/131C12Q2521/107
    • Die vorliegende Erfindung betrifft ein Verfahren zur Synthese einer cDNA in einer Probe in einer enzymatischen Reaktion, wobei das Verfahren die Schritte umfasst, gleichzeitige Bereitstellung eines ersten Enzyms mit Polyadenylierungsaktivität, eines zweiten Enzyms mit reverser Transkriptaseaktivität, eines Puffers, mindestens eines Ribonukleotids, mindestens eines Desoxyribonukleotids, eines Anker Oligonukleotids, Zugabe einer Probe umfassend eine Ribonukleinsäure sowie Inkubation der Agenzien der vorhergehenden Schritte bei einem oder mehreren Temperaturschritten, welche so gewählt sind, dass das erste und das zweite Enzym Aktivität zeigen, wobei zusätzlich in dem selben Reaktionsgemisch eine Amplifikation erfolgt. Die Erfindung betrifft weiter einer Reaktionsgemisch umfassend ein erstes Enzym mit Polyadenylierungsaktivität, eines zweites Enzym mit reverser Transkriptaseaktivität, optional einen Puffer, optional mindestens ein Ribonukleotid, optional mindestens ein Desoxyribonukleotid, optional Anker Oligonukleotid und ein Enzym mit DNA Sytheseaktivität.
    • 本发明涉及一种用于cDNA在酶反应的样品中的合成的方法,所述方法包括同时提供与多聚腺苷酸化的第一酶的步骤,具有逆转录酶活性的第二酶,缓冲液,其包含至少一个核糖核苷酸,至少一种脱氧核糖核苷酸 ,锚寡核苷酸,添加了对一个或多个温度的步骤之前的步骤,其被选择为使得所述第一和所述第二酶表现出活性,此外,放大发生在相同的反应混合物的试剂,其包括核糖核酸的样品以及温育。 本发明还涉及包含多聚腺苷酸化与,具有逆转录酶活性的第二酶,任选的缓冲液中的第一种酶的反应混合物中,任选的至少一种核糖核苷酸,任选地至少一种脱氧核糖核苷酸,任选的锚寡核苷酸和与DNASytheseaktivität的酶。