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

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.

SUBMITTER: Suzuki M

PROVIDER: S-EPMC5614472 | biostudies-literature | 2016 Dec

REPOSITORIES: biostudies-literature

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

Suzuki Masashi M Hayashi Hiroaki H Mizuki Toru T Maekawa Toru T Morimoto Hisao H

Biochemistry and biophysics reports 20161015

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 ...[more]

PMID: 28955977

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Project description:To propose an alternative method of thermoacoustic wave generation based on heating of magnetic nanoparticles (MNPs) using alternating magnetic field (AMF).The feasibility of thermoacoustic wave generation from MNPs by applying a short-burst of AMF or a frequency-modulated AMF is theoretically analyzed. As the relaxation of MNPs is strongly dependent upon the amplitude and frequency of AMF, either an amplitude modulated, fixed frequency AMF (termed time-domain AMF) or a frequency modulated, constant amplitude AMF (termed frequency-domain AMF) will result in time-varying heat dissipation from MNPs, which has the potential to generate thermoacoustic waves. Following Rosensweig's model of specific power loss of MNPs in a steady-state AMF, the time-resolved heat dissipations of MNPs of superparamagnetic size when exposed to a short bursting of AMF and∕or to a linearly frequency chirped AMF are derived, and the resulted acoustic propagation is presented. Based on experimentally measured temperature-rise characteristics of a superparamagnetic iron-oxide nanoparticle (SPION) matrix in a steady-state AMF of various frequencies, the heat dissipations of the SPION under time-domain and frequency-domain AMF configurations that could have practical utility for thermoacoustic wave generation are estimated.The initial rates of the temperature-rise of the SPION matrix were measured at an iron-weight concentration of 0.8 mg∕ml and an AMF frequency of 88.8 kHz to 1.105 MHz. The measured initial rates of temperature-rise were modeled by Rosensweig's theory, and projected to 10 MHz AMF frequency, at which a 1 μs bursting corresponding to a 1.55 mm axial resolution of acoustic detection could contain 10 complete cycles of AMF oscillation and the power dissipation is approximately 84 times of that at 1 MHz. Exposing the SPION matrix to a 1 μs bursting of AMF at 10 MHz frequency and 100 Oe field intensity would produce a volumetric heat dissipation of 7.7 μJ∕cm(3) over the microsecond duration of the AMF burst. If the SPION matrix is exposed to a 1 ms long AMF train at 100 Oe field intensity that chirps linearly from 1 to 10 MHz, the volumetric heat dissipation produced over each 2π phase change of the AMF oscillation is estimated to increase from 0.15 to 1.1 μJ∕cm(3) within the millisecond duration of the chirping of AMF.The heat dissipations upon SPION (∼1 mg∕ml iron-weight concentration) by a 1 μs bursting of 100 Oe AMF at 10 MHz and a 1 ms train of 100 Oe AMF that chirps linearly from 1 to 10 MHz were estimated to determine the potential of thermal-acoustic wave generation. Although thermoacoustic wave generation from MNPs by time- or frequency-domain AMF applications is predicted, the experimental generation of such a wave remains challenging.

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

Publications

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

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