License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: May 13, 2023
Last Modified: May 13, 2023
Protocol Integer ID: 81823
Abstract
tissue dounce homogenisation
Protocol materials
nano-WCatalog ##2018-5ML
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SPEC No:
OPERATOR:
Dounce Homogenisation
Dounce Homogenisation
This method is often used for scabs, warts, or chunks of tissue such as liver. Place ~ 5mm x 5mm piece of scab or tissue into glass homogeniser tube.
Insert teflon plunger into cordless drill, set drill on max speed.
Add equal volume TC water to tube, homogenise(20 strokes) inside Class II BSC cabinet.
Clarify resultant solution at 13000 rpm in Eppendorf centrifuge for 00:01:00
1m
Stand for 00:05:00 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.
5m
Conventional
Conventional
11m
11m
Adsorb 10 µL sample to grid 00:10:00 , inspect to ensure sample does not dry out.
10m
Drain excess sample from grid using filter paper, leave wet.