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通过射频交变磁场选择性加热DNA连接酶实现高效DNA连接

Efficient DNA ligation by selective heating of DNA ligase with a radio frequency alternating magnetic field.

作者信息

Suzuki Masashi, Hayashi Hiroaki, Mizuki Toru, Maekawa Toru, Morimoto Hisao

机构信息

Graduate School of Interdisciplinary New Science, Toyo University, 2100, Kujirai, Kawagoe, Saitama 350-8585, Japan.

Bio-Nano Electronics Research Center, Toyo University, 2100, Kujirai, Kawagoe, Saitama 350-8585, Japan.

出版信息

Biochem Biophys Rep. 2016 Oct 15;8:360-364. doi: 10.1016/j.bbrep.2016.10.006. eCollection 2016 Dec.

Abstract

We present a simple method for efficient DNA ligation utilizing the heat generation of ferromagnetic particles subjected to an ac magnetic field. We carry out the ligation of DNA fragments with cohesive ends using T4 DNA ligase immobilized on the surface of ferromagnetic particles. When a radio frequency alternating magnetic field is applied, ferromagnetic particles dissipate heat and DNA ligase on the particles is selectively heated up and activated with little influence on the annealing of DNA ends, as a result of which the ligation efficiency increases. We show that the ligation efficiency increases with an increase in the field amplitude.

摘要

我们提出了一种利用交变磁场作用下铁磁颗粒产生的热量进行高效DNA连接的简单方法。我们使用固定在铁磁颗粒表面的T4 DNA连接酶对具有粘性末端的DNA片段进行连接。当施加射频交变磁场时,铁磁颗粒会散热,颗粒上的DNA连接酶被选择性加热并激活,而对DNA末端的退火影响很小,其结果是连接效率提高。我们表明,连接效率随场强的增加而提高。

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