Magnetic driven separation techniques - DNA isolation from probiotic food samples

abstrakt

angličtina

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 groups were used for the DNA isolation from crude cell lysates of food products such as yoghurts, probiotic dairy drinks and instant complete initial food. 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, DNA isolation, probiotic food samples, PCR

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

28. 5. 2013

Institute of Immunology and Experimental Therapy Polish Academy of Science

Kudowa Zdrój, Polsko

978-83-928488-3-7

Microbiology and Immunology of Mucosa Probiotics Conference 2013

1

1

27

27

1