Publication detail

The evaluation of magnetic polymethacrylate-based microspheres used for solid phase DNA micro-extraction

TRACHTOVÁ, Š. ZAPLETALOVÁ, H. ŠPANOVÁ, A. HORÁK, D. KOLÁŘOVÁ, H. RITTICH, B.

Original Title

The evaluation of magnetic polymethacrylate-based microspheres used for solid phase DNA micro-extraction

Type

journal article in Web of Science

Language

English

Original Abstract

Using magnetic particles as a solid-phase extraction system is the most frequently used micro-technique for DNA isolation. Particles with a complete covering of magnetic cores by a polymer are hence preferred. Quantitative polymerase chain reaction (qPCR) was used for the evaluation of the polymer coating efficiency of hydrophilic magnetic poly(2-hydroxyethyl methacrylate-co-glycidyl methacrylate) (P(HEMA-co-GMA)) and poly(glycidyl methacrylate) (PGMA) microspheres with/without carboxyl groups. The inhibition effect of magnetic microspheres was identified by the shift in Cq values (ΔCq) after the addition of different amounts of microspheres to PCR mixtures. With the increase of microsphere concentrations, the shift in Cq values to higher values was usually observed. P(HEMA-co-GMA) microspheres containing carboxyl groups extinguished the fluorescence at concentrations over 2 mg mL−1 in a PCR mixture without any influence on the synthesis of PCR products. No PCR products (inhibition of DNA amplification) were detected in the presence of more than 0.8 mg mL−1 in the PCR mixture of PGMA microspheres. Atomic force microscopy (AFM) was used for the determination of the surface morphology of the microspheres. The microspheres were spherical, and their surface was non-porous.

Keywords

magnetic microspheres; quantitative polymerase chain reaction; inhibitory effect; atomic force microscopy

Authors

TRACHTOVÁ, Š.; ZAPLETALOVÁ, H.; ŠPANOVÁ, A.; HORÁK, D.; KOLÁŘOVÁ, H.; RITTICH, B.

RIV year

2015

Released

2. 4. 2015

Publisher

MDPI

Location

Basel, Switzerland

ISBN

2227-9075

Periodical

Chromatography

Year of study

2

Number

2

State

Swiss Confederation

Pages from

156

Pages to

166

Pages count

11

URL

http://www.mdpi.com/2227-9075/2/2/156

Full text in the Digital Library

http://hdl.handle.net/11012/188980

BibTex

@article{BUT133348,
  author="Štěpánka {Trachtová} and Hana {Zapletalová} and Alena {Španová} and Daniel {Horák} and Hana {Kolářová} and Bohuslav {Rittich}",
  title="The evaluation of magnetic polymethacrylate-based microspheres used for solid phase DNA micro-extraction",
  journal="Chromatography",
  year="2015",
  volume="2",
  number="2",
  pages="156--166",
  doi="10.3390/chromatography2020156",
  issn="2227-9075",
  url="http://www.mdpi.com/2227-9075/2/2/156"
}