PCR-FREE LIBRARY PREPARATION USING DOUBLE-STRANDED SPLINT ADAPTORS AND METHODS OF USE

    公开(公告)号:US20240011022A1

    公开(公告)日:2024-01-11

    申请号:US18347440

    申请日:2023-07-05

    IPC分类号: C12N15/10

    CPC分类号: C12N15/1065

    摘要: The present disclosure provides compositions comprising nucleic acid double-stranded splint adaptors, including kits, and methods that employ the double-stranded splint adaptors, e.g., PCR-free workflows. The double-stranded splint adaptors (200) can be used in a one-pot, multi-enzyme reaction to introduce one or more new adaptor sequences into a library molecule. The double-stranded splint adaptor (200) comprises a first splint strand (long splint strand (300)) and a second splint strand (short splint strand (400)), where the first and second splint strands are hybridized together to form the double-stranded splint adaptor (200) having a double-stranded region and two flanking single-stranded regions. The second splint strand (400) carries the new adaptor sequence(s) to be introduced, such as for example a universal binding sequence and/or an index sequence.

    Methods and reagents for nucleic acid analysis

    公开(公告)号:US11781185B2

    公开(公告)日:2023-10-10

    申请号:US17947984

    申请日:2022-09-19

    CPC分类号: C12Q1/6874 C12Q1/6806

    摘要: Provided herein are fluorescently-labeled nucleotide conjugates for nucleic acid analysis. Also provided are reagents used for forming binding complexes between a fluorescently-labeled nucleotide conjugate and a target nucleic acid sequence in the presence of one or more reagents disclosed herein. Binding complexes can be detected in the presence of the one or more reagents. For example, the one or more reagents may contain a photobleaching reducing agent configured to reduce photobleaching resulting from use of the fluorescently-labeled nucleotide conjugate to form the binding complex in a nucleic acid analysis. Such nucleic acid analysis may be used to identify sites of nucleobase binding or incorporation between the target nucleic acid sequence and one or more nucleotide moieties of the fluorescently-labeled nucleotide conjugate in a nucleic acid sequence reaction.