I performed lysozyme assay on my bacteria culture following a suggestion from a paper I read. However, the result that I obtained from the result published is totally different. The result from the paper I read shown a steady decrements of the OD reading. But mine shown a steady increment until they are steady as if they reached the stationary phase of cell growth. Can anyone help me with this?
Dear Mohd Nasharudin,
In lysozyme assays, it is necessary to use dead bacteria. Usually Micrococcus lysodeikticus is used. The use of live bacteria could promote an effect like that you reported.
Avoid long incubation periods. Contamination with live bacteria in long incubation periods would promote increase in absorbance.
Another important thing is to shake the sample before each reading. The best approach is to use a plate reader in kinetic mode with automatic shaking before each reading.
Regards,
Nelder
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