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

摘要

A method for the microextraction of DNA from raw vegetables and highly processed foods ofrnplant origin suitable for PCR analysis was developed. It is based on non-selective binding of DNArnin the presence of PEG 6,000/NaCl to hydrophilic magnetic nonporous poly(2-hydroxyethyl methacrylate-rnco-glycidyl methacrylate) - P(HEMA-co-GMA) microspheres covered by carboxyl groups.rnThe quantitative polymerase chain reaction (qPCR) by in-house designed primers targeting a highlyrnrepetitive rDNA locus allowed for detection of plant DNAs in a wide range of concentrationsrn(0.1 pg/μL to 10 ng/μL). The described procedure is fast and simple. We further demonstrate thatrnrelatively mild acidic treatment of vegetable at elevated temperatures resulted in a dramatic reductionrnof PCR efficiency indicating extensive degradation of DNA during pickling. The describedrnmethod is suitable for the analysis of highly degraded DNA from pickled food products.