Optimisation of Lysis of Bacteria Occuring in Cheeses

abstrakt

angličtina

Presence of Clostridium bacteria in milk is a technological problem for cheese making, as they cause late-blowing defect. Therefore, it is important to rapidly detect these bacteria in milk, e.g. by molecular biotechnological methods like PCR. The aim of this work was to optimise the cell lysis of bacteria of genus Clostridium to receive high concentration of PCR-ready DNA. Lysis solution composed of 10 mM EDTA, 10 mM Tris-HCl and lysozyme (final concentration 3 mg/ml) was considered as the best combination. Recommended incubation times were 1 hour at laboratory temperature with lysozyme and overnight at 55 C with proteinase K. When the lysis was performed with immobilized lysozyme, lower amount of DNA was obtained. The quality of the isolated DNA was confirmed using genus- and species-specific PCRs.

cell lysis, PCR-ready DNA, Clostridium tyrobutyricum

ÜRGEOVÁ, B.; ŠPANOVÁ, A.; RITTICH, B.

15. 6. 2010

Pamida International Ltd.

Košice, Slovensko

978-80-970168-4-5

International Scientific Conference on Probiotics and Prebiotics, Conference Proceedings

159

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