Storage of virus at -80 - (Feb/26/2007 )
Currently we store a flavivirus at -80 in borate saline. However there is a problem with stability and so I was wondering if anyone had suggestions of how else to store the virus.
We use borate saline as this is what is used in the heamagluttination assay the virus is required for (positive control). We have found that we cannot store it in culture media as this contains FBS which in turn interferes with the HA assay.
Thanks,
P
we store Lentivirus in -80C in media but one can store them in PBS as well. We used to store AAV also in PBS in -80C.
I am not sure abt flavi virus.
What sort of length of time do you manage to store them for (and maintain viability) in PBS?
we store them (lentivirus) indefinitely in -80C. But every freeze thaw cycle reduces the virus's infectivity and we dont use them after 3rd freeze thaw cycle.
We store all kinds of virusses @ -80°C (rota-virusses, RSV, HIV...) until needed for analysis or other experiments (which can take years).
It's true though that freeze-thawing reduces infectivity (most virusses have membranes and they can be destroyed by water crystals).
It's true though that freeze-thawing reduces infectivity (most virusses have membranes and they can be destroyed by water crystals).
See our problem isn't even one of a freeze/thaw in that we are only doing one cycle before the aliquots are used in the assay. However after a few months at -80 the ability of the virus to agglutinate the RBCs drops to virtually nothing over time. I'm wondering if it is a problem specifically due to the heamagluttinin then and that the virus is still present/capable of infecting cells. Easy enough to find out I suppose.
It's true though that freeze-thawing reduces infectivity (most virusses have membranes and they can be destroyed by water crystals).
See our problem isn't even one of a freeze/thaw in that we are only doing one cycle before the aliquots are used in the assay. However after a few months at -80 the ability of the virus to agglutinate the RBCs drops to virtually nothing over time. I'm wondering if it is a problem specifically due to the heamagluttinin then and that the virus is still present/capable of infecting cells. Easy enough to find out I suppose.
Some viruses cannot be frozen once they are thawed like adenovirus. They simply dont work. Once thawed use the whole aliquot even if you have to keep them in 4C for 24 more hours.
It's true though that freeze-thawing reduces infectivity (most virusses have membranes and they can be destroyed by water crystals).
See our problem isn't even one of a freeze/thaw in that we are only doing one cycle before the aliquots are used in the assay. However after a few months at -80 the ability of the virus to agglutinate the RBCs drops to virtually nothing over time. I'm wondering if it is a problem specifically due to the heamagluttinin then and that the virus is still present/capable of infecting cells. Easy enough to find out I suppose.
Some viruses cannot be frozen once they are thawed like adenovirus. They simply dont work. Once thawed use the whole aliquot even if you have to keep them in 4C for 24 more hours.
That's what I was meaning. I do use the whole aliquot I would never re-freeze it. The aliqouts are all frozen at the same time then used one aliqout per assay - anything remaining is discarded. The problem appears after thawing a fresh aliqout which has remained at -80 for a few months. It's a pain
