Detail publikace

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

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

Originální název

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

Typ

článek v časopise ve Web of Science, Jimp

Jazyk

angličtina

Originální abstrakt

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.

Klíčová slova

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

Autoři

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

Vydáno

1. 11. 2017

ISSN

1940-087X

Periodikum

Journal of visualized experiments : JoVE

Ročník

129

Číslo

1

Stát

Spojené státy americké

Strany od

1

Strany do

9

Strany počet

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"
}