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    • 9. 发明申请
    • NUCLEIC ACID ISOLATION
    • 核酸分离
    • WO1997034909A1
    • 1997-09-25
    • PCT/FR1997000496
    • 1997-03-20
    • BIO MERIEUXCROS, PhilippeELAISSARI, AbdelhamidMABILAT, ClaudePICHOT, ChristianRODRIGUE, MarcSANTORO, Lise
    • BIO MERIEUX
    • C07H01/08
    • C12N15/101C12N15/1006C12Q1/6806
    • A method for aqueous phase nucleic acid isolation from a sample, comprising a step of nucleic acid adsorption on a particulate substrate, is disclosed. The method comprises an adsorption reagent preparation step (a) providing an adsorption reagent that includes a sol consisting of an aqueous continuous phase and a dispersed particulate substrate phase including a functionalised particulate polymer prepared by polymerising (1) a first water-soluble acrylamide or acrylamide derivative monomer, (2) at least one cross-linking agent and (3) at least one second water-soluble, cationic and functional monomer, said polymer having a predetermined lower critical solubility temperature (LCST) of 25-45 DEG C; a contact step (b) wherein the adsorption reagent is contacted with the sample containing the nucleic acid; an adsorption step (c) wherein, to carry out the contact step (b), at least one parameter is selected for the reaction medium, said parameters being a pH no higher than 7, an ionic strength no higher than 10 M, and a temperature lower than the polymer LCST; a separation step (d) wherein the dispersed phase is separated from the continuous phase, optionally after it has been observed that adsorption has occurred; and a desorption step (e) wherein the nucleic acid is desorbed from the particulate substrate by increasing the ionic strength until an ionic strength higher than 10 M is achieved.
    • 公开了一种从样品中分离水相核酸的方法,包括在颗粒基质上的核酸吸附步骤。 该方法包括一种吸附试剂制备步骤(a),该步骤(a)提供一种吸附试剂,该吸附试剂包括由含水连续相组成的溶胶和分散的颗粒基质相,所述分散的颗粒底物相包括通过聚合制备的官能化颗粒聚合物(1)第一水溶性丙烯酰胺或丙烯酰胺 衍生单体,(2)至少一种交联剂和(3)至少一种第二水溶性阳离子和功能性单体,所述聚合物具有25-45℃的预定的较低临界溶解温度(LCST); 接触步骤(b),其中所述吸附剂与含有所述核酸的样品接触; 吸附步骤(c)其中,为了进行接触步骤(b),为反应介质选择至少一个参数,所述参数为不高于7的pH,不高于10 -2的离子强度 M,温度低于聚合物LCST; 分离步骤(d),其中分散相与连续相分离,任选地在已经观察到发生吸附之后; 和解吸步骤(e),其中通过增加离子强度直到达到高于10μM的离子强度,从核粒子脱离。