Magnetic driven separation techniques

lecture

English

Rapid and sophisticated nucleic acids isolation strategies play very important role in molecular diagnostics of microorganisms. Magnetic driven separation techniques using magnetic solid particles are one of modern methods to speed up and facilitate the nucleic acids (DNA and RNA) separation and purification procedures. The problem of falsely negative results in PCR analysis of real food samples due to the presence of extracellular PCR inhibitors can be solved using solid phase systems adsorbing genomic DNA. In our work various types of magnetic non-porous poly(2-hydroxyethyl methacrylate-co-glycidyl methacrylate) - P(HEMA-co-GMA) microspheres, poly(2-hydroxyethyl methacrylate-co-etylene dimethacrylate) - P(HEMA-co-EDMA) and poly(glycidylmethacrylate) - (PGMA) covered by different content of carboxyl groups1 were used for the DNA isolation from crude cell lysates of food products such as yoghurts, probiotic dairy drinks and instant complete initial food. These types of food products represent a matrix characterized by a complex composition including the presence of inhibitors that affect the course of PCR2. It was shown that tested microspheres are suitable for the PCR and qPCR ready DNA isolations from these types of food samples. The amount of isolated DNA correlated with the content of carboxyl groups on the microsphere surface. Higher quantity of DNA was isolated using microspheres with higher content of carboxyl groups. Isolated DNAs were without PCR inhibitors which were demonstrated using PCR and real-time PCR. Only one PCR product was amplified in real-time PCR using DNA isolated by tested microspheres from all dairy products.

magnetic particles, probiotic food products, PCR

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

30. 5. 2013

Institute of Immunology and experimental therapy Polish Academy of Science

Kudowa Zdrój, Polsko

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