Isolation of Mosquito-Associated Virus from Mosquito Homogenate

Streszczenie

Source: Huang, D., et al. Mosquito-Associated Virus Isolation from Field-Collected Mosquitoes. J. Vis. Exp. (2022).

This video demonstrates the step-by-step procedure for the in vitro isolation of mosquito-associated viruses from mosquito homogenates using fibroblast-like mammalian cells.

Protokół

1. Virus isolation
   NOTE: All steps were carried out in a biosafety level 2 (BSL-2) laboratory. The safety level requirement of the biosafety laboratory was determined by the biosafety risk assessment based on the regulations of different countries and regions. The process must be performed in a biosafety cabinet.
   1. Remove the cell culture medium from the 24-well plates and add 100 µL of the cell culture maintenance medium and 100 µL of the supernatant of the mosquito homogenate (P0) to each well.
   2. Incubate the plates at 28 °C or 37 °C for 60 min, gently shaking them every 15 min to pr....

Materiały

Lista materiałów użytych w tym artykule

Nazwa	Firma	Numer katalogowy	Komentarze
24-well plates	CORNING	3524	Containers for cell
Screw cap storage tubes (2 mL)	biofil	FCT010005
Refrigerator (-80 °C)	sanyo	MDF-U54V
Incubator (37 °C)	Panasonic	MCO-18AC	Instrument for cell culture
Dulbecco’s minimal essential medium (DMEM)	Gibco	C11995500BT	medium for vertebrate cell lines