stability of TOPO vector at RT - (Feb/06/2006 )
I have a less-usual question. For some reason, Invitorgen sent my TOPO kit order "Saturday delivery". No one was around, and the package sat at room temp until Monday - it thawed completely. I expect the competent cells are dead, but I'm wondering if the vector is still useable. Anyone ever tried using vector that had sat out that long?
(Invitrogen is replacing the order, but I'm curious)
Thanks!
-Rebecca
whilst I haven't tested it, my feeling is it wont work. I know that TOPO kits can be quite sensitive and lose activity over time. The topoisomerase is connected to the vector, and if its been out at room temperature, chances are its not active anymore. There is no way then to get your PCR product ligated.
I doubt it will work.
Even casual use over the course of five to six months (taking it out of the freezer, etc.) is generally too much.
Try it once.
I'd call invitrogen to see what you might arrange.
-Matt
Thanks for the suggestions. Out of curiousity, I tried using the thawed vector side-by-side with the replacement sent by Invitrogen, which had not been thawed. They both worked, equally well!
So, while I certainly wouldn't advise letting your vector sit around a RT, if it happens the vector may well survive 
If your budget is tight, it is certainly worth checking before tossing the vector.
-Rebecca