Vapor Phase Seed Sterilization

ASOP-0002 Version 1

Table of Contents

1.0: Purpose

The purpose of this SOP is to provide instructions on how to use chlorine gas to surface sterilize Arabidopsis thaliana seeds.  This is a more thorough sterilization than ASOP-0001: Ethanol Seed Sterilization that can be used for seeds that are especially susceptible to contamination. 

2.0: Safety

Standard laboratory personal protective equipment (PPE) should be worn when executing this technique.  Refer to Material Safety Data Sheets (SDS) for hazards and appropriate handling precautions.

3.0: Definitions and Acronyms

  • ASOP: Astrobotany Standard Operating Procedure
  • A. thaliana: Shorthand for Arabidopsis thaliana
  • HCl: Shorthand for concentrated hydrochloric acid

4.0: Materials

  • Concentrated Hydrochloric Acid
  • Bleach
  • 1.5 mL microcentrifuge tubes
  • Container that can be closed with a lid (e.g. plastic bin) 
  • Pipette capable of delivering 3 mL of HCl
  • 250 mL glass beaker

5.0: Equipment

  • Fume Hood or Biosafety Cabinet (BSC)

6.0: Procedure

  1. 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.
  2. Place your tubes inside a container that can be closed with a lid.
  3. Place that container inside a fume hood.
  4. Make sure the microcentrifuge tube lids are open.
  5. Place a 250 mL beaker inside the container with the tubes.
  6. Put 100 mL of bleach in the beaker.
  7. Pipette 3 mL of concentrated HCl into the bleach.
  8. Immediately seal the lid of the container after this addition.
  9. Let the seeds sit for 3-5 hours.
  10. Dispose of all waste material properly.

7.0: References

The original protocol is attributed to Maud Hinchee (Monsanto), Steven Clough (UI-Urbana-Champaign), and Andrew Bent (UI-Urbana-Champaign).

Version History

  • Version 1 | Description: ASOP-0002 page created. | Effective: 16Jun2023
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