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    METHOD FOR ISOLATING NON-VESICULAR MIRNA
    4.
    发明公开
    METHOD FOR ISOLATING NON-VESICULAR MIRNA 审中-公开

    公开(公告)号：US20240287498A1

    公开(公告)日：2024-08-29

    申请号：US18570344

    申请日：2022-06-15

    申请人： QIAGEN GmbH

    发明人： Martin SCHLUMPBERGER

    IPC分类号： C12N15/10

    CPC分类号： C12N15/1013

    摘要： The present invention provides methods and kits for isolating cell-free, non-vesicular miRNAs from a biological sample, such as serum and plasma, by using an acidic binding buffer comprising a buffering agent to promote binding of non-vesicular miRNAs to a solid phase comprising anion exchange groups. Furthermore, binding of non-vesicular miRNAs to the solid phase may be improved due to the presence of a crowding agent and by increasing the surface charge density of the solid phase.

    ENRICHMENT METHOD
    6.
    发明申请
    ENRICHMENT METHOD 有权

    公开(公告)号：US20230028205A1

    公开(公告)日：2023-01-26

    申请号：US17785478

    申请日：2020-12-16

    申请人： QIAGEN GmbH

    发明人： Martin SCHLUMPBERGER ,  Karolin SPITZER

    IPC分类号： C12N15/10

    摘要： A method is provided for enriching extracellular DNA from a biological sample comprising extracellular DNA and extracellular vesicles, wherein the method comprises: (a) preparing a binding mixture comprising—the biological sample, —a solid phase comprising anion exchange groups, —an acidic binding buffer comprising a buffering agent, and binding extracellular DNA to the solid phase comprising anion exchange groups; (b) separating the solid phase with the bound extracellular DNA from the remaining binding mixture, wherein the remaining binding mixture comprises extracellular vesicles. The method may furthermore comprise processing the remaining binding mixture to enrich one or more biological targets of interest therefrom, wherein processing may comprise (c) enriching as biological targets extracellular vesicles and/or extracellular RNA from the remaining binding mixture.

    METHOD FOR SEPARATING NUCLEIC ACID MOLECULES BY SIZE
    7.
    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.

    POLYMERASE ENZYME
    10.
    发明申请
    POLYMERASE ENZYME 有权

    公开(公告)号：US20210254032A1

    公开(公告)日：2021-08-19

    申请号：US17181787

    申请日：2021-02-22

    申请人： QIAGEN GMBH

    发明人： Ralf Peist ,  Frank Narz ,  Nicole Grasse ,  Anne Schieren ,  Thea Rutjes

    IPC分类号： C12N9/12

    摘要： The present invention relates to a family B polymerase enzyme comprising the following mutations, a mutation in the motif A region, wherein a threonine is the second amino acid of the motif A region, wherein the motif A region is homologous to amino acids 408 to 410 in 9° N, a mutation leading to a reduction or elimination of the 3′-5′ exonuclease activity if present in said family B polymerase, wherein said polymerase additionally comprises one or both of the following mutations in the motif A region, L408Y or L408F and/or P410S or P410G. It may be used in DNA sequencing and may be present in a kit.