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Liquid phage release assay
| Link to this post | posted today, 14:35 | |
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Hello, My question is regarding the expected titer of phage in the supernatant of a liquid phage release assay from a potential lysogen. I'm trying to verify my potential G. terrae phage lysogens using liquid phage release assay. After 2 rounds of streak, I set up liquid culture for these potential lysogens for 3 days. I then collected and titrated the supernatant using spot test. The titers are at least 1x10^5 pfu/ml with clearing and plaque patterns consistent with the serial dilutions. I'm wondering if a titer of 1x10^5 pfu/ml is sufficient for me to conclude that I have a lysogen that is releasing phages, or if I should expect higher titers to draw that conclusion. Is there a recommended threshold of titer for this assay? Thanks! Ping |
| Link to this post | posted today, 14:39 | |
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Ping, I think a titer of any number is sufficient if you have purified the bacteria such that residual phage from somewhere outside of the cell is removed. I would not evaluate this is a vacuum. are the cells homo-immune to that phage infection. Build the evidence and then see what it tells you. We do not have empirical numbers, but you can collect that data. (And then we would for each phage.) Best, debbie |
| Link to this post | posted today, 14:46 | |
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Thanks for the quick response! Debbie. I'll move on to the next step with the homo-immunity test and will keep you posted with more data and most likely, more questions. Ping |