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Abstract

DNA of five pathogenic bacterial strains (Salmonella typhimurium 51 K61 K, Campylobacter jejuni CCUG 11284, Escherichia coli 0157, Listeria monocytogenes 4B NCTC and Yersinia enterocolitica Y79) were isolated and purified by our partners in the framework of Validation and standardisation of diagnostic polymerase chain reaction for detection of food-borne pathogens (Food PCR) project (PL QLRT-1999-00226).

Our aim was to produce certified reference material from the received samples that are stable for a long time and transportable on room temperature. We set up stability and recovery tests for studying the stability of purified DNA. The DNA was tested in the case of different storage methods: in TAE buffer, adsorbed on different media (paper, ionic exchanger resins, glass, activated carbon) and dried on the wall of different vials (glass, polypropylene tubes).

The effect of different storage temperature ( -20, 5, 20, 40, 100 degrees Celsius) on the stability was studied as well. Recovery of dried DNA was the highest (85-100%) from the wall of polypropylene tubes. It was 60-80% from different papers and below 10% from ionic exchanger resins.

Significant difference was found in the stability of dried DNA applying the polypropylene tubes of different suppliers. The supplier of applied 0.5 ml volume polypropylene tubes was selected on the basis of DNA degradation and recovery results. The produced genomic DNA reference materials were produced on room temperature by vacuum drying. The prepared reference materials contain 1mirco-grams purified DNA in a dried form, which amount is enough for performing 20-100 PCR tests. ests.

Additional information

Authors: PROKISCH J, EC-JRC, Institute for Reference Materials and Measurements, Geel (BE);TRAPMANN S, EC-JRC, Institute for Reference Materials and Measurements, Geel (BE);CATALANI P, EC-JRC, Institute for Reference Materials and Measurements, Geel (BE);SCHIMMEL H, EC-JRC, Institute for Reference Materials and Measurements, Geel (BE);PAUWELS J, EC-JRC, Institute for Reference Materials and Measurements, Geel (BE)
Bibliographic Reference: An oral report given at: 2nd General Meeting of the FOOD-PCR Project. Held in: Lund (SE) 17-19 June, 2001
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