Hello, I had purchased MS2 (freeze-dried) from ATCC: 15597-B1 (https://www.atcc.org/products/15597-b1). I revived the culture according to the method given in the product sheet. For propagation, I have used E. coli BL21 instead of the recommended strain. Also, the revival was done past the 30 day time period that was mentioned. I have separated the E.coli by centrifugation at 15000 rpm. Is it possible that the MS2 remained in the E. coli cell? Or is it that I stored the freeze-dried culture for too long before reviving it? Or is it because I have used the wrong strain of E. coli? I still have half o the free dried pellet but before using it I want to make sure that the method I am following is correct.
Is there some other way apart from plaque assay and PCR to ensure that the MS2 is present in the filtrate?
MS2 is a male-specific phage that adsorbs to sex pili, such as those encoded by the F episome. BL21 does not harbour such episome (it is F-), thus it is not able to propagate MS2. I would try again infecting with an F+ strain.
@Pieree Beguin. I tried reviving the MS2 culture with a wild type E.coli that we had. I still could not see any plaques. So does this mean the wild type E. coli does not harbour the F episome?
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