Publication detail

DNA-magnetic particle binding analysis by dynamic and electrophoretic light scattering

HADDAD, Y. DOSTÁLOVÁ, S. KUDR, J. ZÍTKA, O. HEGER, Z. ADAM, V.

Original Title

DNA-magnetic particle binding analysis by dynamic and electrophoretic light scattering

Type

journal article in Web of Science

Language

English

Original Abstract

Isolation of DNA using magnetic particles is a field of high importance in biotechnology and molecular biology research. This protocol describes the evaluation of DNA-magnetic particles binding via dynamic light scattering (DLS) and electrophoretic light scattering (ELS). Analysis by DLS provides valuable information on the physicochemical properties of particles including particle size, polydispersity, and zeta potential. The latter describes the surface charge of the particle which plays major role in electrostatic binding of materials such as DNA. Here, a comparative analysis exploits three chemical modifications of nanoparticles and microparticles and their effects on DNA binding and elution. Chemical modifications by branched polyethylenimine, tetraethyl orthosilicate and (3-aminopropyl) triethoxysilane are investigated. Since DNA exhibits a negative charge, it is expected that zeta potential of particle surface will decrease upon binding of DNA. Forming of clusters should also affect particle size. In order to investigate the efficiency of these particles in isolation and elution of DNA, the particles are mixed with DNA in low pH (similar to 6), high ionic strength and dehydration environment. Particles are washed on magnet and then DNA is eluted by Tris-HCl buffer (pH = 8). DNA copy number is estimated using quantitative polymerase chain reaction (PCR). Zeta potential, particle size, polydispersity and quantitative PCR data are evaluated and compared. DLS is an insightful and supporting method of analysis that adds a new perspective to the process of screening of particles for DNA isolation.

Keywords

Chemistry; Issue 129; DNA isolation; magnetic particle; chemical modification; dynamic light scattering; quantitative PCR

Authors

HADDAD, Y.; DOSTÁLOVÁ, S.; KUDR, J.; ZÍTKA, O.; HEGER, Z.; ADAM, V.

Released

1. 11. 2017

ISBN

1940-087X

Periodical

Journal of visualized experiments : JoVE

Year of study

129

Number

1

State

United States of America

Pages from

1

Pages to

9

Pages count

9

BibTex

@article{BUT143912,
  author="Yazan Abdulmajeed Eyadh {Haddad} and Simona {Dostálová} and Jiří {Kudr} and Ondřej {Zítka} and Zbyněk {Heger} and Vojtěch {Adam}",
  title="DNA-magnetic particle binding analysis by dynamic and electrophoretic light scattering",
  journal="Journal of visualized experiments : JoVE",
  year="2017",
  volume="129",
  number="1",
  pages="1--9",
  doi="10.3791/56815",
  issn="1940-087X"
}