高比活度α-[32P]dATP和α-[32P]ATP的快速、简便且改良的制备方法。
The rapid, simple and improved preparation of high specific activity alpha-[32P]dATP and alpha-[32P]ATP.
作者信息
Symons R H
出版信息
Nucleic Acids Res. 1977 Dec;4(12):4347-55. doi: 10.1093/nar/4.12.4347.
Abstract
An improved method is described for the rapid and simple preparation of alpha-[32P]dATP and alpha-[32P]ATP from 32Pi in good yields and with specific activities from 20 - 150 Ci/mmol. The two-step procedure involves the chemical synthesis of the mononucleotide followed by its enzymic conversion to the triphosphate with myokinase (EC 2.7.4.3) and pyruvate kinase (EC 2.7.1.40) in the presence of trace amounts of dATP or ATP to prime the reaction. The two steps are carried out in the same reaction flask and the only purification step required is a step-wise elution from a column of DEAE-cellulose.
摘要
描述了一种改进的方法，可从³²Pi快速简便地制备α-[³²P]dATP和α-[³²P]ATP，产率高，比活度为20 - 150 Ci/mmol。两步法包括单核苷酸的化学合成，然后在微量dATP或ATP存在下，用肌激酶（EC 2.7.4.3）和丙酮酸激酶（EC 2.7.1.40）将其酶促转化为三磷酸酯以引发反应。这两步在同一反应烧瓶中进行，唯一需要的纯化步骤是从DEAE-纤维素柱上进行分步洗脱。
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