Study on the encapsulation of magnetic cores using real time polymerase chain reaction

abstrakt

angličtina

Solid phase magnetic driven separation techniques in combination with molecular diagnostic methods (polymerase chain reaction, PCR) have become popular to speed up and facilitate the nucleic acids separation and purification procedures. Magnetic particles are mostly based on magnetic iron oxide (magnetic core) covered by polymer surfaces suitable for modification by different functional groups (e.g. –COOH, -NH2). Free or incomplete encapsulated cores and some components applied in the preparation of magnetic particles can interfere with polymerase chain reaction [1, 2]. Therefore perfectly encapsulation of magnetic cores and removing all potential PCR inhibiting components are key factors for the application of magnetic particles in molecular diagnostics. For this reasons, polymerase chain reaction in quantitative real time (qPCR) and the software supplied with the qPCR, were used for the estimation of the inhibitory effect of different types of magnetic microparticles. The set of magnetic non-porous poly(2-hydroxyethyl methacrylate-co-glycidyl methacrylate) - P(HEMA-co-GMA) microspheres (8 types, (Figure 1)) and poly(glycidylmethacrylate) - P(GMA) (6 types, (Figure 1)) covered by different content of carboxyl groups and 3 types of compounds used for magnetic core encapsulation were studied. Linear regression analyses were used for the evaluation of the influence of the particles or compounds on the qPCR course. From results, it follows that the tested particles can be split up to 3 groups according to the effect on the DNA amplification. The results presented in this report show that polymerase chain reaction in real time is suitable method for the evaluation of the quality of magnetic particles.

magnetic particles, PCR magnetic cores encapsulation

TRACHTOVÁ, Š.; ŠPANOVÁ, A.; RITTICH, B.; HORÁK, D.

10. 6. 2014

Technische Universitat Dresden

Dresden, Germany

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