In the User’s Laboratory:

1. Empty the contents of used 15 or 50ml tubes ​that have only been used for the tissue culture. Ensure these tubes have never been used for viral, bacterial, or yeast work. Note: Only Greiner Bio-one tubes or Eppendorf Tubes® BioBased are suitable for this process. Just empty the contents of the tubes (e.g., in Virkon) without rinsing and securely close the lids.

2. Continue collecting these tubes in racks or suitable containers only from the tissue culture.

Washing up Facility:

1. Ensure the tubes are empty and with closed lids. Only Greiner Bio-one tubes or Eppendorf Tubes® BioBased should be collected for this process.

2. Erase any permanent marker pen labels from the tubes using 75%-100% ethanol.

3. Remove lids from the tubes. Fully submerge both tubes and lids in either 1% Distel (Scientific Lab Supplies part number: TRI1366, 5L) or 1% Chemgene HLD4L Concentrate (Clear Unfragranced, 5 Litre XTM309-C STARLAB (UK) Ltd) for 30 minutes to 1 hour. Use distilled water to dilute the disinfectant to 1%, and always prepare fresh disinfectant for each session.

4. Thoroughly wash the tubes and lids with tap water. Continue washing until there's no froth when shaking the tubes with water.

5. Once washed, rinse the tubes and lids with distilled water. Dry the tubes and lids completely in a drier.

6. Pack an equivalent number of dry tubes and their corresponding lids into an autoclave bag (either 15, 50ml tubes with 15 lids or 40, 15ml tubes with 40 lids).

7. Seal the bag using autoclave tape and label the outside specifying whether they are 50ml or 15ml tubes and proceed to autoclave.

8. After autoclaving, either move the unopened bags to a trolley in a communal area for collection or deliver the bags directly to users.

 

Back in the User’s Laboratory:

1. Retrieve the bags and open the bag in a Class II cabinet (Tissue culture hood).

2. Individually pair each tube with its corresponding lid and return them to the bag for use in the tissue culture room only.

3. If any lids don't fit their tubes securely, dispose of those tubes in a regular trash bin and not in the autoclave bin since these tubes are uncontaminated.

4. Tubes sterilized through this method are deemed suitable for all tissue culture tasks. Nevertheless, a pilot run for your specific tasks is advisable.

You can wash and reuse these tubes multiple times until they deform or their lids no longer secure properly.

1. Open the lids of the tip boxes (or aluminium boxes or bags) inside Class II cabinets (Tissue culture hood).

2. Close each tube with its corresponding lid and return them to the box, drawer, or autoclave bags for reuse.

3. Tubes treated with this protocol are deemed sterile for all non-cell culture work (e.g., cloning, protein work, western blotting, solution preparation). However, they're not suitable for tissue culture or RNA work. These tubes can be reused and washed multiple times until they deform or their lids no longer fit securely. These tubes can be reused and washed multiple times until they deform or their lids no longer fit securely.