HI,
I am working on vesicular stomatits virus ( VSV) and i should harvest high titer of VSV for about 500 ml. up to now i did TCID50 assay to measure the vsv titer i harvested and i got 10-5 . But i need 10-7 titer.
for inoculation of vero cell line by VSV, after three time washing the vero flask by PBS, i add 1 ml VSV and 5 ml DMEM to T25 flask of 80%-90% confluency for about 1 hour and after that put out the virus and finally i add DMEM with pen/strep and 1% FBS and incubate for 24 hour to see CPE.
Hello, if you need 10-7 from 10-5 its easy step to receive by dilution, but I see you wrote wrong titer I think you have 10^5 but need 10^7 without minus.
In this case you need ultra centrifugation for concentration of your sample and increasing titer.
See the reference for more info.
Good luck.
Hello, if you need 10-7 from 10-5 its easy step to receive by dilution, but I see you wrote wrong titer I think you have 10^5 but need 10^7 without minus.
In this case you need ultra centrifugation for concentration of your sample and increasing titer.
See the reference for more info.
Good luck.
Karen A. Darbinyan
thank you karen for your answer.
Let me explain more about the titer which i got. after i harvest the vsv, i did TCID50 for getting the titer of virus which could make CPE in 50% of 96-well microplate.
so i made serial dilution of my vsv stock from 10 to the power of -1 (10^ -1) to 10 to the power of -12 ( 10^ -12). after 24 hour incubation, i checked my microplate and its CPE. I realized that i had CPE from wells of 10^ -1 ( which was more concentrated) to 10^ -5 ( which was less concentrated than ten to the power of -1).
so i mean how can i have a high titer VSV which can make CPE even in its lower titer? I mean even it can make CPE in the wells with dilution of 10^ -7 .
Also ultra centrifugation will be effective in this case?
Thank you very much.
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