baculovirus lost titer

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baculovirus lost titer

Postby volodya_tx » Oct 06 2017 9:11 pm

Dear Colleagues,

My baculoviral P3 stock stopped working for protein expression (storage time ~5 months at 4 C).
I tried to re-amplify from P2->P3 virus and P3->P4 viruses (also stored for 5 months at 4 C), my cells got infected and then I harvested supernatant 120 h post-infection (cell viability was ~30%), but when these viruses were used for expression they did not work again.
I feel I should start all over again from bacmid transfection?
volodya_tx
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Re: baculovirus lost titer

Postby Anorak » Feb 09 2018 2:47 pm

volodya_tx wrote:Dear Colleagues,

My baculoviral P3 stock stopped working for protein expression (storage time ~5 months at 4 C).
I tried to re-amplify from P2->P3 virus and P3->P4 viruses (also stored for 5 months at 4 C), my cells got infected and then I harvested supernatant 120 h post-infection (cell viability was ~30%), but when these viruses were used for expression they did not work again.
I feel I should start all over again from bacmid transfection?


Hey, It might be a long shot, but I have similar problem at the moment and I don't really know what to do? Have you found some emergency solution or you just started over?
Anorak
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