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公开(公告)号:US20210102189A1
公开(公告)日:2021-04-08
申请号:US16650598
申请日:2017-09-27
申请人: QIAGEN GmbH
IPC分类号: C12N15/10
摘要: A phenol-free method for isolating a nucleic acid from a sample is provided, said method comprising the following steps: a) adding a precipitation buffer to a sample to prepare an acidic precipitation mixture wherein said precipitation buffer comprises a metal cation precipitant and a buffering agent, has a pH value of 4.0 or less and does not comprise an organic solvent selected from aprotic polar solvents and protic solvents and wherein the acidic precipitation mixture comprises the metal cation precipitant in a concentration of less than 200 mM and precipitating proteins; b) separating the precipitate from the supernatant, wherein the supernatant comprises small RNA having a length of less than 200 nt and large RNA having a length of at least 1000 nt; and c) isolating a nucleic acid from the supernatant. The present method allows to avoid the use of organic solvents during protein precipitation. Also provided is a precipitation buffer.
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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.
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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
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.
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公开(公告)号:US20230357747A1
公开(公告)日:2023-11-09
申请号:US18343296
申请日:2023-06-28
申请人: QIAGEN GmbH
IPC分类号: C12N15/10
CPC分类号: C12N15/1003
摘要: A phenol-free method for isolating a nucleic acid from a sample is provided, said method comprising the following steps: a) adding a precipitation buffer to a sample to prepare an acidic precipitation mixture wherein said precipitation buffer comprises a metal cation precipitant and a buffering agent, has a pH value of 4.0 or less and does not comprise an organic solvent selected from aprotic polar solvents and protic solvents and wherein the acidic precipitation mixture comprises the metal cation precipitant in a concentration of less than 200 mM and precipitating proteins; b) separating the precipitate from the supernatant, wherein the supernatant comprises small RNA having a length of less than 200 nt and large RNA having a length of at least 1000 nt; and c) isolating a nucleic acid from the supernatant. The present method allows to avoid the use of organic solvents during protein precipitation. Also provided is a precipitation buffer.
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公开(公告)号:US20230009972A1
公开(公告)日:2023-01-12
申请号:US17785251
申请日:2020-12-16
申请人: QIAGEN GmbH
IPC分类号: C12N15/10
摘要: The present invention pertains to methods and kits for enriching extracellular nucleic acids such as vesicular RNA from a sample comprising extracellular vesicles. Accordingly to the methods an acidic binding mixture is prepared comprising the sample and anion exchange particles and binding extracellular vesicles to the anion exchange particles. After separating the anion exchange particles comprising the bound extracellular vesicles from the remaining mixture, bound extracellular vesicles are lysing the in the presence of at least one detergent and released RNA is bound to the anion exchange particles. The anion exchange particles with the bound RNA from the lysate are then eluted.
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