NCEM Drop - Cell Pellet Dounce Homogenisation (TM-014)

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SPEC No:

OPERATOR:

Remove cells from the flasks by scraping with a disposable plastic scraper and poor into a 10mL centrifuge tube.

Pellet the cells in a bench top centrifuge at 2000rpm,0h 0m 0s 0h 2m 0s , remove supernatant.

Resuspend the cells in equal volume TC water and transfer to glass homogeniser tube.

Insert teflon plunger into cordless drill, set drill on max speed.

Homogenise(20 strokes) inside Class II BSC cabinet.

Clarify resultant solution at 13000rpm,0h 0m 0s in Eppendorf centrifuge for 0h 1m 0s

Stand for 0h 5m 0s to permit viruses to diffuse back into solution, from the debris, and provide an interface for the sampling of membrane associated viruses. Use supernatant for sample below.

Adsorb 10µL sample to grid 0h 10m 0s , inspect to ensure sample does not dry out.

Drain excess sample from grid using filter paper, leave wet.

Stain 0h 1m 0s

Drain & dry using filter paper