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# NCEM Drop - Cell Pellet Dounce Homogenisation (TM-014) V.2

DOI

dx.doi.org/10.17504/protocols.io.bp2l69j4dlqe/v2

sandra.crameri¹

¹CSIRO ACDP

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sandra.crameri

OPEN ACCESS

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DOI: dx.doi.org/10.17504/protocols.io.bp2l69j4dlqe/v2

**Protocol Citation:** sandra.crameri 2023. NCEM Drop - Cell Pellet Dounce Homogenisation (TM-014). protocols.io

https://dx.doi.org/10.17504/protocols.io.bp2l69j4dlqe/v2Version created by sandra.crameri

**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:** 81825

## Abstract

cell pellet dounce homogenisation

## Protocol materials

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## HEADER

1. SAN:

SPEC No:

OPERATOR:

## Dounce Homogenisation

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

3. Pellet the cells in a bench top centrifuge at **2000 rpm**, remove supernatant.

2m

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

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

6. Homogenise (20 strokes) inside Class II BSC cabinet.

7. Clarify resultant solution at **13000 rpm** in Eppendorf centrifuge for **00:01:00**

1m
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8 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

11m

9 Adsorb 🗥 10 µL sample to grid 🕒 00:10:00 , inspect to ensure sample does not dry out.

10m

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

11 Stain 🌐 nano-W Catalog ##2018-5ML 🕒 00:01:00

1m

12 Drain & dry using filter paper