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

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

WoS Article

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.

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.

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

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

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

2017

01.04.2015

ELSEVIER

Spojené království Velké Británie a Severního Irska

0960-3085

FOOD AND BIOPRODUCTS PROCESSING

94

1

United Kingdom of Great Britain and Northern Ireland

375

381

7