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1.发明申请Method for the Selective Enrichment of Double-Stranded Dna from Nucleic Acid Mixtures 审中-公开从核酸混合物选择性富集双链Dna的方法公开(公告)号：US20100009434A1
公开(公告)日：2010-01-14
申请号：US11991947
申请日：2006-09-13
CPC分类号： C12N15/1003
摘要： The invention relates to a method for stripping undesired nucleic acid components from double-stranded DNA, in particular, super-coiled plasmid DNA. The method according to the invention is characterised by the steps: (a) provision of a mixture containing completely and/or partly double-stranded nucleic acids and optionally single-stranded nucleic acids; (b) resuspension of the mixture from step (a) in an aqueous, low-molarity buffer system with low ionic strength and low buffer effect; (c) adjusting conditions in the mixture from step (b), under which the completely and/or partly double-stranded nucleic acids are denatured; (d) further addition of buffer and a polymer component to the mixture from step (c); (e) incubation of the mixture from step (d) for a time which is sufficient for the formation of an aqueous two-phase system with an upper and lower phase; and (f) removal of the upper phase containing the single-strand nucleic acid and collection of the double-strand nucleic acid from the lower phase.
摘要翻译： 本发明涉及一种从双链DNA，特别是超螺旋质粒DNA中去除不想要的核酸成分的方法。 根据本发明的方法的特征在于以下步骤：（a）提供含有完全和/或部分双链核酸和任选的单链核酸的混合物; （b）将混合物从步骤（a）重悬于具有低离子强度和低缓冲效应的低摩尔浓度缓冲体系中; （c）调节来自步骤（b）的混合物中的完全和/或部分双链核酸变性的条件; （d）进一步向步骤（c）的混合物中加入缓冲剂和聚合物组分; （e）将来自步骤（d）的混合物温育足以形成具有上部和下部相的水相两相体系的时间; 和（f）除去含有单链核酸的上层和从下相收集双链核酸。
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2.发明授权公开(公告)号：US08034562B2
公开(公告)日：2011-10-11
申请号：US10558326
申请日：2004-05-28
IPC分类号： C12Q1/68
CPC分类号： C12N15/1003
摘要： The invention relates to a process for isolation of plasmid DNA from biomass by means of an aqueous 2-phase system having a polymer component and a salt component, characterized in that the resuspension of the biomass employed, the alkaline lysis of the biomass, the neutralization of the alkaline lysis batch and the separation of the plasmid DNA from the contaminants are carried out in a single reaction vessel (one-pot process) rendered possible in that the neutralization of the alkaline lysis batch is carried out in one and the same container by addition of potassium phosphate and one component of the aqueous 2-phase system is therefore already present. The second component of the aqueous 2-phase system is a PEG having a molecular weight of the mathematical average of about 600 g/mol to 1,000 g/mol.
摘要翻译： 本发明涉及通过具有聚合物组分和盐组分的水相二相体系从生物质中分离质粒DNA的方法，其特征在于所用生物质的再悬浮，生物质的碱裂解，中和 的碱性裂解批料和质粒DNA与污染物的分离在单个反应容器（一锅法）中进行，使得可能的碱裂解批料的中和在同一容器中通过 因此，已经存在磷酸钾的添加和两相水相的一种组分。 水相二相体系的第二组分是数均分子量为约600g / mol至1,000g / mol的PEG。
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3.发明申请METHOD FOR SELECTIVELY AND REVERSIBLY ADSORBING NUCLEIC ACIDS ON A CARRIER MATERIAL 审中-公开用于选择性和反向吸附载体材料上的核酸的方法公开(公告)号：US20100036109A1
公开(公告)日：2010-02-11
申请号：US12084667
申请日：2006-11-07
IPC分类号： C07H21/00
CPC分类号： C12N15/1006
摘要： The invention relates to a process for selective, reversible adsorption of nucleic acids to a support material, which process is characterized in that (a) a nucleic acid-containing mixture having a cosmotropic salt content of at least 1 mole is contacted with said support material, whereupon the nucleic acid(s) is (are) adsorbed from said nucleic acid-containing mixture to said support material, and (b) the nucleic acid(s) bound to the support material is (are) detached from the support material by means of an aqueous solution having a salt content of no more than 1 mole.
摘要翻译： 本发明涉及将核酸选择性地，可逆地吸附到载体材料上的方法，该方法的特征在于：（a）具有至少1摩尔的共成盐含量的含核酸的混合物与所述载体材料接触 因此，所述核酸从所述含核酸混合物吸附到所述载体材料上，并且（b）与载体材料结合的核酸通过 含盐量不超过1摩尔的水溶液的方法。
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4.发明申请公开(公告)号：US20060286080A1
公开(公告)日：2006-12-21
申请号：US10558326
申请日：2004-05-28
CPC分类号： C12N15/1003
摘要： The invention relates to a process for isolation of plasmid DNA from biomass by means of an aqueous 2-phase system having a polymer component and a salt component, characterized in that the resuspension of the biomass employed, the alkaline lysis of the biomass, the neutralization of the alkaline lysis batch and the separation of the plasmid DNA from the contaminants (such as e.g. cell debris, RNA and gDNA) are carried out in a single reaction vessel (one-pot process). According to the invention, this is rendered possible in that the neutralization of the alkaline lysis batch is carried out in one and the same container by addition of potassium phosphate and one component of the aqueous 2-phase system is therefore already present, and in that the second component of the aqueous 2-phase system is a PEG having a molecular weight of the mathematical average of about 600 g/mol to 1,000 g/mol, but is preferably formed from a mixture of PEG 600 and PEG 1000.
摘要翻译： 本发明涉及通过具有聚合物组分和盐组分的水相二相体系从生物质中分离质粒DNA的方法，其特征在于所用生物质的再悬浮，生物质的碱裂解，中和 的碱性裂解批料和质粒DNA与污染物（例如细胞碎片，RNA和gDNA）的分离在单个反应容器（一锅法）中进行。 根据本发明，可以通过加入磷酸钾在一个容器中进行碱裂解批料的中和，因此已经存在一个水相二相体系的一个组分，因此， 水相二相体系的第二组分是数均分子量为约600g / mol至1,000g / mol的PEG，但优选由PEG 600和PEG 1000的混合物形成。
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