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Titration of JEV - Plaque assay for Japanese encephalitis virus (Sep/06/2006 )



I am doing apoptosis study on japanese encephalitis virus,
for initial titration i am using BHK-21[ 370C and 5% C02]cell lines,
2 overlay media i used already Bac plaque agarose and Carboxy methyl cellulose,

what the problem is , after two days my cells undergone stress and all cell got round up even in control also,

i m using 5% FBS in Overlay, and 2% FBS in MEM for virus diluent,

Please tell me the protocol for this assay using Plaque agarose,


what is your final concentration of agarose in the overlay......i bring mine ti 1 % final concentration - different virus and cell line though. for BHK u could try final conc of 1% tragacanth gum, 2% FCS also.....eliminates problem of accidently heat damaging cells