- 专利标题: Method for Library Preparation in Next Generation Sequencing by Enzymatic DNA Fragmentation
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申请号: US18274541申请日: 2022-01-28
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公开(公告)号: US20240076803A1公开(公告)日: 2024-03-07
- 发明人: Matthias Wahl
- 申请人: Miltenyi Biotec B.V. & Co. KG
- 申请人地址: DE Bergisch Gladbach
- 专利权人: Miltenyi Biotec B.V. & Co. KG
- 当前专利权人: Miltenyi Biotec B.V. & Co. KG
- 当前专利权人地址: DE Bergisch Gladbach
- 优先权: EP 154220.4 2021.01.29
- 国际申请: PCT/EP2022/051979 2022.01.28
- 进入国家日期: 2023-07-27
- 主分类号: C40B50/06
- IPC分类号: C40B50/06
摘要:
The invention is directed to a method for obtaining a nucleic acid library of a sample comprising polynucleotides comprising the steps a. multiplying the polynucleotides by a polymerase b. fragmenting the multiplied polynucleotides by creating nicks c. coupling an oligonucleotide sequence to the nicks to create the target library wherein step a) is performed by providing A, T, G, C and U nucleotides wherein the molar ratio of T and U is between 200:1 and 5:1 and step b) is performed by excision of the U nucleotides. characterized in that after step b), the nicks are provided with a polymerase exhibiting 5′ #3′ exonuclease activity, thereby filling in the 3′ recessing ends and removing the 5′ overhangs of the nicks.
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