I am testing phosphate stress on a rhizobia species. I will then be doing enzyme assays on the cultures.
Is it fine to grow one culture and then repeat the assay in triplicate. Or is it necessary to culture the bacteria in triplicate and then assay each repeat in triplicate?
if you need a biological replication you have to perform cultivation in triplicates. Otherwise it will be just an assay replication, what is not you want to check if I correctly understand.
if you need a biological replication you have to perform cultivation in triplicates. Otherwise it will be just an assay replication, what is not you want to check if I correctly understand.
1. You are testing phosphate stress on Rhizobia species, means it belong to biological functioning of Rhizobia towards phosphate stress. So it is better to grow the culture in triplicates.
2. Later on, you are checking the enzyme assay of the cultures. To avoid any mis-handling, environmental effect or any other factor, it is suggested to perform enzyme assay in triplicates, so that you may get most probably accurate results.
If you want to check the reproducibility of the assay, then there is no need of triplicate cultures.
Growing a culture in three flasks originating from single overnight culture does not count as biological replicate. Thus you'd need to grow separate overnight cultures (from separate colonies on plate, if you're using them) or to grow it at another time.
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