DNA amplification by real-time polymerase chain reaction (RT-PCR) was used for the evaluation of efficiency
of polymer coating of magnetic hydrophilic 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 on real-time polymerase chain reaction (RT-PCR) course was evaluated by
regression analysis after the addition of different concentrations of tested microspheres to PCR mixtures. Microspheres
mostly did not interfere in RT-PCR till the concentration 50 µg/25µl PCR mixture. No relationship between
Fe content (and microsphere diameter) and inhibition effect was found. Microspheres containing carboxyl
groups extinguished the fluorescence at lower concentrations (10-20 µg/25µl PCR mixture) without inhibition of DNA amplification as
PCR products were detected using agarose gel electrophoresis. Negative effect of maghemite on PCR course was partially reduced by
coating of magnetic core by silica or polymers. Two inhibition mechanisms of DNA amplification were discussed in this work.
Keywords: Magnetic microspheres, inhibitory effect, real-time polymerase chain reaction, regression analysis.
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