I have identified and characterized the new phage I have found some months ago, and I was able to capture the electron microscopy pictures and also sequence it. But I have had a serious issue maintaining the phage titer. In fact, and most of the time, the phage titer decreases one to two logs in a matter of days.
Have you seen such a phenomenon? I already have been working with other phages but the decrease in titer has not been this drastic.
I have tried the plate lysate method and the conventional liquid amplification method.
Thank you for sharing your opinion.
- If you are storing the phages along with chloroform, your phages might be chloroform sensitive, and the pfu is going down.
- Sometimes vortexing the phage suspension causes the breaking of phage capsid and tails leading to phage inactivation.
- Store phage suspension at -80 for maintaining a longer time.
Yes. Its very common with some phages. It depend in phage and also how did you prepared the stock. In general using gelatine to the concentration 2% will stabilise the stock and keep you phages safe for longer. For more stable phages keep environmental sample for 1-3 days in refrigerator and than use it for phage isolation. This will select for more stable phages.
If you want to keep your phages in 4 degree Celsius, it's better if you store them in SM buffer (50mM Tris-HCl pH7. 5, 100mM NaCl, 8mM MgSO4, 0.01% Gelatin (v/w)) instead of in LB or ddH2O. I observed when I made phage dilution with water and keep it in 4 degree, the titer will decrease quickly.
For maintaining titer a longer time you can keep in -80 in glycerol.
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