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and was fully characterized (accession number KX197192.1).Following isolation, the virus was passed once on Ae. Viral stocks were then produced in VERO cells and stored at −80 °C until use.This mixture was incubated at room temperature for 10 min and then centrifuged at 12000g for 10 min at 4 °C.The supernatant was removed, and the RNA pellet was washed with 300 μL of 75% ethanol.Seven- to ten-day-old female mosquitoes were starved for 18 h prior to artificial feeding.
Both assays included a control group fed on uninfected cultured cells mixed with defibrinated rabbit blood.
Tissues were individually transferred to a 1.5-m L DNAse/RNAse-free microtube containing 300 μL of mosquito diluent and were stored at −80 °C until further usage.