I am unable to store marine bacteriophages for a long time with an SM buffer at -4 C°, -20 C° and -80 C°. Please share the correct protocol for this.
How long do you consider 'a long time'? I'm most familiar with phage being kept long-term (for a year or more) at -80 in 50% glycerol and phage buffer. Some old papers (including this 1962 one: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1057894/) suggest that 4 degrees should work fine as well, but only when starting with a high-titer lysate.
I suggest you to read this book chapter. Note that the conditions that work well for one phage might not work for your phage. We routinely keep our cleared and filter-sterilized phage lysates at 4 degree, but some phage lysates tend to degrade faster than others for unknown reasons. You can freeze your lysate at -80 degree in 50% glycerol. Hope this will help you.
Chapter Phage Production and Maintenance of Stocks, Including Expect...
Hi,
If you are storing the virus stocks/lysates at -4 degree C (stocks i use have a high titer e.g. 1e9 or more), the virus are viable for a month or 1.5 months. It is better to refresh the stocks every month while storing in 4 degrees. I have used this technique and it works as the viruses i study are highly stable.It is better to make a routine to determine the virus infectivity in between the storage time to make sure you dont loose them.
And as described above, 50 % glycerol stock is best when storing the virus for a year or more at -80 degrees. We dont store virus lysates at -20.
Regards
Bina
Honestly, I've never had trouble with infectivity of my sterile-filtered cyanophage stocks stored in TM buffer with a little drop of chloroform at 4 degrees C for four years.
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