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Recent Activity
Growing Liquid Cultures for lysogens
| Link to this post | posted today, 18:12 | |
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Greetings, I am trying to grow liquid cultures of lysogens to be able to complete the sensitivity assay (11.5). However, when trying to grow up liquid cultures, there is very little bacterial growth, even though I have growth on a patch plate. I am concerned that I will not be able to get a lawn on the petri dishes. Does anyone have any advice? Thanks, Beth |
| Link to this post | posted 58 minutes ago | |
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Hi Beth, Have you colony purified the lysogen? You want to remove exogenous phage from the bacteria that you are growing in a liquid culture. If you are starting from a plate seeded (or loaded) with phage, you will get lots of killing in a liquid culture. Best, debbie |
| Link to this post | posted 53 minutes ago | |
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Hi Debbie, Thank you for your reply. I streaked for single colonies and then did three patch plates. Would it be advisable to streak more? Best, Beth |
| Link to this post | posted 12 minutes ago | |
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Beth, Hmm. If you original colony was surrounded by phage, you may be picking up exogenous phage. So streak it 3 times on agar. Then grow a culture and plate. Is the background filled with plaques? You can perform a streak of that bacteria on a plate of wild type bacteria, but use the culture to make your lysogen plate for testing. If that is what you have done, then your lysogen is very leaky…. You may never get a culture grown, but if you do and you plate, and it is a webbed looking plate. sometimes the spot titers will still show up, even in the mess. Have you tried to grow up the lysogen using a phage seeded plate (instead of cultures the cells from a spot). if so, do you have %lysogeny numbers? Just all something to think about. debbie |