Eastern equine encephalitis virus original isolate. This Eastern equine encephalitis virus[EEE 2090-EEE 2090] is preserved under Viral Storage Medium -80C.Tests for the presence of mycoplasmae were negative. To confirm its identity the virus has been partly sequenced.
Product Risk Group:
Viral Storage Medium -80C
Infectivity tested and quantified
Infectivity was proven by plaque titration. Infectious titre was determined to be 4x 10^7 PFU/ml.
Production cell line:
Passaged 32 times on suckling mice in 1993 and stored freeze-dried. New stock prepared by one passage on Vero E6 cells on 18th November 2010.
RT-PCR followed by sequencing of the amplified product
We recommend to grow the virus on VeroE6 cells cultivated in MEM Eagle with Earle"s BSS (BioWhittaker, 12-136F). Following additives should be added to 500 ml of medium: 5ml Nonesential amino acids (Biochrom AG, K0293), 5 ml Sodium Pyruvate 100mM, (Biochrom AG, L0473), 5 ml L-Alanyl-L-glutamine 200mM (Biochrom AG, K0302), 5 ml Penicilin/ Streptomycin/ Amphotericin (BioWhittaker 17-745E), 25 ml FBS heat inact.s.American (Invitrogen, 10108-165). Multiplicity of infection (MOI) of 0,00001 can be used for the infection and the infectious medium can be harvested 42hours after infection.