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    METHOD FOR SEPARATING NUCLEIC ACID MOLECULES BY SIZE
    1.
    发明申请
    METHOD FOR SEPARATING NUCLEIC ACID MOLECULES BY SIZE 有权

    公开(公告)号：US20220340954A1

    公开(公告)日：2022-10-27

    申请号：US17620615

    申请日：2020-06-26

    申请人： QIAGEN GmbH

    发明人： Lothar BREITKOPF ,  Jasper STROEDER

    IPC分类号： C12Q1/6806 ,  C12N15/10

    摘要： The present invention provides a poly(alkylene oxide) polymer based size selective nucleic acid enrichment method for enriching target nucleic acid molecules from a nucleic acid containing sample which comprises target nucleic acid molecules and non-target nucleic acid molecules, wherein the target nucleic acid molecules are longer than the non-target nucleic acid molecules, the method comprising (a) preparing a binding mixture comprising—the nucleic acid containing sample, —a poly(alkylene oxide) polymer and—a salt and binding nucleic acid molecules to a solid phase which comprises a functional group, preferably carboxylated magnetic particles, wherein the bound nucleic acid molecules comprise target nucleic acid molecules; (b) preferably separating the solid phase with the bound nucleic acid molecules from the remaining sample; (c) contacting the solid phase with the bound nucleic acid molecules at least once with a reagent composition comprising a poly(alkylene oxide) polymer and a salt to selectively elute non-target nucleic acid molecules, wherein the concentration (w/v) of the poly(alkylene oxide) polymer in the reagent composition of step (c) is lower than the concentration (w/v) of the poly(alkylene oxide) polymer in the binding mixture of step (a); (d) optionally washing the bound target nucleic acid molecules; and (e) eluting the bound target nucleic acid molecules from the solid phase. Said method allows the size selective purification of target DNA molecules and is particularly suitable for sequencing applications. It is more time- and cost efficient than prior art methods and provides excellent purification results. Moreover, further methods and kits are provided.

    METHOD FOR ENRICHING NUCLEIC ACIDS BY SIZE
    2.
    发明申请
    METHOD FOR ENRICHING NUCLEIC ACIDS BY SIZE 有权

    公开(公告)号：US20220259637A1

    公开(公告)日：2022-08-18

    申请号：US17620612

    申请日：2020-06-26

    申请人： QIAGEN GmbH

    发明人： Lothar BREITKOPF ,  Jasper STROEDER ,  Markus SPRENGER-HAUSSELS

    IPC分类号： C12Q1/6806 ,  C12Q1/686 ,  C12N15/10 ,  C12N15/82

    摘要： The present invention provides a poly(alkylene oxide) polymer based size selective method for enriching nucleic acid molecules having a length below a cut-off value from a nucleic acid containing sample, the method comprising: (a) preparing a binding mixture comprising —the nucleic acid containing sample, —a poly(alkylene oxide) polymer and —a salt and binding nucleic acid molecules of different sizes to a solid phase which comprises a functional group, preferably carboxylated magnetic particles; (b) separating the solid phase with the bound nucleic acid molecules from the remaining sample; and (c) contacting the solid phase with the bound nucleic acid molecules at least once with an elution composition comprising a poly(alkylene oxide) polymer and a salt to selectively elute nucleic acid molecules having a length below the cut-off value from the solid phase while larger nucleic acid molecules having a length above the cut-off value remain bound to the solid phase, wherein the concentration (w/v) of the poly(alkylene oxide) polymer in the elution composition is lower than the concentration (w/v) of the poly(alkylene oxide) polymer in the binding mixture of (a); (d) separating the solid phase with the bound larger nucleic acid molecules from the eluted nucleic acid molecules; and (e) optionally further purifying the eluted nucleic acid molecules. The method is particularly useful for separating extracellular target nucleic acids by size.