Timeline for Bacterial cells disappearing from culture with time
Current License: CC BY-SA 4.0
7 events
| when toggle format | what | by | license | comment | |
|---|---|---|---|---|---|
| Jan 15 at 13:45 | comment | added | CaroZ | This might be an unrelated question, but may I know what protein you are expressing ? | |
| Jan 11 at 12:48 | comment | added | acvill | Is it possible that your E. coli strain is a lysogen? Prophage induction at high cell density could explain why your culture is crashing | |
| Jan 10 at 16:03 | comment | added | ksdjnf | (end) -Vigilance: you will never remove all the phage, you'll just keep them under control. Make sure everyone watches for signs of lysis and communicates them. | |
| Jan 10 at 16:03 | comment | added | ksdjnf | (cont) -Filter pipet tips may prevent transferring aerosolized phage. -Try to prevent aerosols in general. Be gentle with your cells and don't leave plates open. -You mentioned trying alternative strains, but you might look into this a bit more. Some common phage receptors differ between B and K strains, for instance. -You could try low temperature or cold inductions to potentially reduce induction. -Regular maintenance: once phage has established itself, you probably want to have regularly-scheduled deep cleaning (especially surfaces where it can linger) to keep it down. | |
| Jan 10 at 16:03 | comment | added | ksdjnf | That's tough. I was in a lab that struggled with recurrent phage contamination before, and there isn't a great solution. Fumigation sounds like a good idea - we weren't able to do that. We found a few things seemed to work well enough to keep things under control: this might not work for your combination of phage and cells. -Virkon is better than bleach. Wash all flasks and regularly wipe down counters, pipettes, and common instruments and spaces. -High temperature is better than autoclaving. See if there's an oven you can use to sterilize flasks. | |
| Jan 10 at 7:05 | comment | added | Juno | I thought of that possibility as well. The transformed colonies on plates with antibiotic looks normal. A few times I have observed wierd colony morphology (not quite circular, but distorted). I did try plaque assay, but couldn't notice any area of clearance on LB-agar plate with bacterial lawn. We fumigated (formaldehyde-permanganate) the lab and all equipment, washed all glassware with bleach, and alcohol followed by autoclaving before preparing fresh media. Still the problem persists. Do you know of any other method to detect/ get rid of phage contamination? | |
| Jan 9 at 14:52 | history | answered | ksdjnf | CC BY-SA 4.0 |