We are trying to find a protocol which can serve as a mother protocol for our research regarding dengue. Any help?
I would recommend you use the mosquito cell line for dengue
C6/36
Mosquito; Larva whole
Growth medium: Minimum essential medium (Eagle) with 2mM L-glutamine and Earle’s BSS adjusted to contain 0.35 g/L Na bicarbonate , 0.1 mM non-essential amino acids, Tryptose Phosphate Broth 10% (of 2.9% stock soln.), and 1.0 mM Na pyruvate, 90%; Fetal calf serum, 10%
Temperature: 28 C
Methods of Sub Culturing :
Remove medium add TPVG. After 1-2 minutes remove TPVG. After 5-10 minutes add fresh medium ; agitate to dissociate cells, make homogeneous suspension and seed new culture.
Split Ratio of 1:4 is suggested
Sub Culture Int.--: 4-5 days
Fluid Renewal--: Twice per week
Note : TPVG , PBS with 0.1% Trypsin , 0.2% Versene and 0.05% Glucose
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