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

Trachtová, Štěpánka; Španová, Alena; Tóth, Judit; Prettl, Zsolt; Horák, Daniel; Gyenis, János; Rittich, Bohuslav

doi:10.1016/j.fbp.2014.05.001

Cited by 15 publications

(7 citation statements)

References 38 publications

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“…Copolymers are widely used as ion exchangers [ 16 , 17 , 18 , 19 , 20 ], macromolecular supports for enzyme immobilization [ 21 , 22 , 23 ] or catalysts [ 24 ], column packings for HPLC [ 10 , 11 , 12 , 25 ], and dye remover [ 26 , 27 ]. Microspheres based on GMA are also widely applied in bioseparation [ 28 , 29 ], cell isolation [ 30 ], nucleic acid and protein purification and immunoassay [ 31 , 32 , 33 , 34 ]. Fluorescence and encoded GMA-based microspheres can be used as fluorescent sensor [ 35 ] and material that shows a particularly important role in labeling, detection, and high-throughput screening [ 36 , 37 ].…”

Section: Introductionmentioning

confidence: 99%

Studies on Preparation, Characterization and Application of Porous Functionalized Glycidyl Methacrylate-Based Microspheres

Pączkowski

Gawdzik

2021

Materials

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A one-step swelling and polymerization technique was used in the synthesis of porous glycidyl methacrylate (GMA) and ethylene glycol dimethacrylate (EGDMA) monodisperse polymeric microspheres. The polystyrene (PS) seed obtained in the dispersion polymerization was used as a shape template. The presence of epoxide rings in the chemical structure of microspheres enables their post-polymerization chemical modifications involving: the Diels-Alder reaction with sodium cyclopentadienide and maleic anhydride, the reaction with 4,4′-(bismaleimido)diphenylmethane, and the thiol-Michael reaction with methacryloyl chloride and 2-mercaptopropionic acid. Changing the reaction mixture composition—the amounts of crosslinking monomer and PS seed as well as the type and concentration of porogen porous microspheres of different porous structures were obtained. Their porous structures were characterized in the dry and swollen states. The copolymers obtained from the equimolar monomers mixture modified in the above way were applied as the column packing materials and tested in the reverse-phase HPLC (High-Performance Liquid Chromatography). A few factors influencing morphology and porous structure of microspheres were studied.

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Section: Introductionmentioning

confidence: 99%

Studies on Preparation, Characterization and Application of Porous Functionalized Glycidyl Methacrylate-Based Microspheres

Pączkowski

Gawdzik

2021

Materials

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“…The magnetic hybrid nanocomposites were synthesized to perform simple and fast DNA separation [22]. Hydrophilic non-porous magnetic microspheres containing carboxyl groups were tried for the isolation of DNA from different food matrices and the highest isolation was achieved with the microspheres containing high content of carboxyl groups within the medium containing polyethylene glycol and NaCl [23]. The hydrophobic sorbents or the sorbents with anionic groups lead to the formation of a strong connection between the sorbent and DNA by strong electrostatic attraction or hydrophobic interactions.…”

Section: Introductionmentioning

confidence: 99%

Comparative DNA isolation behaviours of silica and polymer based sorbents in batch fashion: monodisperse silica microspheres with bimodal pore size distribution as a new sorbent for DNA isolation

Günal

Kip

Öğüt

et al. 2017

Artificial Cells, Nanomedicine, and Biotechnology

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Monodisperse silica microspheres with bimodal pore-size distribution were proposed as a high performance sorbent for DNA isolation in batch fashion under equilibrium conditions. The proposed sorbent including both macroporous and mesoporous compartments was synthesized 5.1 μm in-size, by a "staged shape templated hydrolysis and condensation method". Hydrophilic polymer based sorbents were also obtained in the form of monodisperse-macroporous microspheres ca 5.5 μm in size, with different functionalities, by a developed "multi-stage microsuspension copolymerization" technique. The batch DNA isolation performance of proposed material was comparatively investigated using polymer based sorbents with similar morphologies. Among all sorbents tried, the best DNA isolation performance was achieved with the monodisperse silica microspheres with bimodal pore size distribution. The collocation of interconnected mesoporous and macroporous compartments within the monodisperse silica microspheres provided a high surface area and reduced the intraparticular mass transfer resistance and made easier both the adsorption and desorption of DNA. Among the polymer based sorbents, higher DNA isolation yields were achieved with the monodisperse-macroporous polymer microspheres carrying trimethoxysilyl and quaternary ammonium functionalities. However, batch DNA isolation performances of polymer based sorbents were significantly lower with respect to the silica microspheres.

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“…The above mentioned mechanisms were reviewed by Rittich and Španová (). Hydrophilic magnetic nonporous poly(2‐hydroxyethyl methacrylate‐ co ‐glycidyl methacrylate) (P(HEMA‐ co ‐GMA)) and poly(glycidyl methacrylate) (PGMA) microspheres covered with carboxyl groups have been successfully used for the microisolation of high‐molecular‐weight bacterial DNA from different complex matrices containing PCR inhibitors such as fermented dairy products and cheeses (Rittich et al, ; Trachtová et al, ). However, they were not used for DNA isolation from plant samples, yet.…”

Section: Introductionmentioning

confidence: 99%

Application of magnetic polymethacrylate-based microspheres for the isolation of DNA from raw vegetables and processed foods of plant origin

Trojánek

Kovařı́k

Španová

et al. 2017

J Food Process Preserv

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A method for the microextraction of DNA from raw vegetables and highly processed foods of plant origin suitable for PCR analysis was developed. It is based on non-selective binding of DNA in the presence of PEG 6,000/NaCl to hydrophilic magnetic nonporous poly(2-hydroxyethyl methacrylate-co-glycidyl methacrylate) -P(HEMA-co-GMA) microspheres covered by carboxyl groups.The quantitative polymerase chain reaction (qPCR) by in-house designed primers targeting a highly repetitive rDNA locus allowed for detection of plant DNAs in a wide range of concentrations (0.1 pg/lL to 10 ng/lL). The described procedure is fast and simple. We further demonstrate that relatively mild acidic treatment of vegetable at elevated temperatures resulted in a dramatic reduction of PCR efficiency indicating extensive degradation of DNA during pickling. The described method is suitable for the analysis of highly degraded DNA from pickled food products. Practical applicationsDNA-based methods, such as the polymerase chain reaction (PCR), represent an important genetic approach for identification of plant species composition of foods. DNA from processed foods varies in both quality and quantity between the protocols used. Plant samples are generally characterised by a complex composition containing various inhibitors of PCR. Current methods have been tailored for specific samples while no universal protocol is available. Development of a simple universal procedure leading to PCR-ready DNA would be beneficial. Isolation strategies based on solid phase systems have become popular for PCR-ready DNA isolations from complex matrices.Here we applied magnetic hydrophilic P(HEMA-co-GMA) microspheres to extract DNA from raw vegetables and highly processed foods of plant origin. Proposed small scale PCR-ready DNA extraction protocol was comparable with a silica-based DNeasy Plant Mini Kit (Qiagen) and classical CTAB extraction methods in quality and amplificability of DNA while it is less costly and time consuming.

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

Cited by 15 publications

References 38 publications

Studies on Preparation, Characterization and Application of Porous Functionalized Glycidyl Methacrylate-Based Microspheres

Studies on Preparation, Characterization and Application of Porous Functionalized Glycidyl Methacrylate-Based Microspheres

Comparative DNA isolation behaviours of silica and polymer based sorbents in batch fashion: monodisperse silica microspheres with bimodal pore size distribution as a new sorbent for DNA isolation

Application of magnetic polymethacrylate-based microspheres for the isolation of DNA from raw vegetables and processed foods of plant origin

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