WARNING: This protocol involves chlorine gas and should only be performed by professional researchers with biosafety training in a laboratory environment. DO NOT ATTEMPT WITHOUT PROPER TRAINING AND EQUIPMENT.
This protocol uses chlorine gas to surface sterilize Arabidopsis thaliana seeds. This is for when you want to perform a more thorough sterilization than an ethanol wash.
The original protocol is attributed to Maud Hinchee (Monsanto), Steven Clough (UI-Urbana-Champaign), and Andrew Bent (UI-Urbana-Champaign).
Wear gloves and use proper ventilation.
- Make sure the microcentrifuge tubes your seeds are in are properly labeled. Pencil and paper works the best as chlorine gas can sometimes wipe sharpie clean off of a surface.
- Place your tubes inside a container that can be closed with a lid.
- Place that container inside a fume hood.
- Make sure the tubes are open. Centrifuge tube lids off.
- Place a 250 mL beaker inside the container with the tubes.
- Put 100 mL of bleach in the beaker.
- Pipette 3 mL of concentrated HCl into the bleach.
- Immediately seal the lid of the container after this addition.
- Let the seeds sit for 3-5 hours.
- Dispose of all waste material properly.