Publication detail

Solid-phase DNA isolation from food matrices using hydrophilic magnetic microspheres

TRACHTOVÁ, Š. ŠPANOVÁ, A. TÓTH, J. PRETTL, Z. HORÁK, D. GYENIS, J. RITTICH, B.

Original Title

Solid-phase DNA isolation from food matrices using hydrophilic magnetic microspheres

Type

journal article in Web of Science

Language

English

Original Abstract

Dynamic light scattering (DLS) was used to monitor the coil–globule transition of calf thymus and bacterial DNAs by poly(ethylene glycol) (PEG 600 and PEG 6,000) in aqueous NaCl solutions. The contribution of PEG 600 and PEG 6,000 to DNA coil diameter change was investigated. Compaction of DNA molecules was observed for a PEG 6,000 concentration ranging from 5.5 to 8 %. Hydrophilic non-porous poly(2-hydroxyethyl methacrylate-co-glycidyl methacrylate) - P(HEMA-co-GMA), poly(2-hydroxyethyl methacrylate-co-ethylene dimethacrylate) - P(HEMA-co-EDMA), and poly(glycidyl methacrylate) (PGMA) microspheres containing carboxyl groups were used for DNA isolation. The highest DNA yield was achieved using 16 % PEG 6,000 and 2.0 M NaCl. The amount of isolated DNA correlated with the content of carboxyl groups on the microsphere surface. RNA adsorption on the surface of the microspheres tested was also studied. RNA recovery was more than one order lower than DNA recovery under the same conditions. All types of microspheres were tested for DNA isolation from crude cell lysates of different dairy products in solutions of 16 % PEG 6,000 and 2.0 M NaCl. Isolated DNAs were without PCR inhibitors, which was demonstrated using real-time PCR including melting analysis of the amplicons.

Keywords

DNA compaction; Magnetic microspheres; DNA isolation; RNA; Polymerase chain reaction (PCR)

Authors

TRACHTOVÁ, Š.; ŠPANOVÁ, A.; TÓTH, J.; PRETTL, Z.; HORÁK, D.; GYENIS, J.; RITTICH, B.

RIV year

2015

Released

1. 4. 2015

Publisher

ELSEVIER

Location

Spojené království Velké Británie a Severního Irska

ISBN

0960-3085

Periodical

FOOD AND BIOPRODUCTS PROCESSING

Year of study

94

Number

1

State

United Kingdom of Great Britain and Northern Ireland

Pages from

375

Pages to

381

Pages count

7

BibTex

@article{BUT126302,
  author="Štěpánka {Trachtová} and Alena {Španová} and Judit {Tóth} and Zsolt {Prettl} and Daniel {Horák} and János {Gyenis} and Bohuslav {Rittich}",
  title="Solid-phase DNA isolation from food matrices using hydrophilic magnetic microspheres",
  journal="FOOD AND BIOPRODUCTS PROCESSING",
  year="2015",
  volume="94",
  number="1",
  pages="375--381",
  doi="10.1016/j.fbp.2014.05.001",
  issn="0960-3085"
}