Copper-Catalyzed Alkyne-Azide Cycloaddition on the Solid Phase for the Preparation of Fully Click-Modified Nucleic Acids.

Copper-Catalyzed Alkyne-Azide Cycloaddition on the Solid Phase for the Preparation of Fully Click-Modified Nucleic Acids.
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铜催化的炔烃叠氮化物固相环加成反应制备完全点击修饰的核酸

DOI:
10.1007/978-1-4939-9216-4_11
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发表时间:
1973
影响因子:
--
通讯作者:
Mayer G
Mayer G
中科院分区:
--
文献类型:
--
作者:
Rosenthal M;Pfeiffer F;Mayer G

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点击化学已经成为一种广泛使用的将修饰插入DNA的方法。由于其商业可用性,5 ' -乙基-脱氧尿嘧啶(EdU)通常被并入DNA中,通过点击反应进行后续修饰。然而,在固相合成过程中的脱保护过程中,它被部分氧化,导致酮不再可以进行点击修饰。为了实现含edu DNA的高保真固相合成,本协议描述了在脱保护前在固相上进行点击反应的程序。然后,按照标准程序对DNA进行去保护和纯化,选择叠氮化物对EdU的完全修饰可以通过HPLC和HPLC/MS进行分析。
Click chemistry has become a widely used method to insert modifications into DNA. Due to its commercial availability, 5′-ethynyl-deoxyuridine (EdU) is commonly incorporated into the DNA for subsequent modification by click reaction. However, it is partially oxidized during deprotection during solid-phase synthesis, resulting in a ketone that is no longer accessible for click modification. To enable the high-fidelity solid-phase synthesis of EdU-containing DNA, this protocol describes a procedure to perform the click reaction on the solid phase before deprotection. Afterwards, the DNA can be deprotected and purified according to standard procedures, and the full modification of EdU with the azide of choice can be analyzed by HPLC and HPLC/MS.
DOI: 10.1021/acs.bioconjchem.5b00668
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影响因子: 4.7
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