Ethanol Seed Sterilization

ASOP-0001 Version 1

Table of Contents

1.0: Purpose

The purpose of this SOP is to provide instructions on how to quickly sterilize seed batches of Arabidopsis thaliana (or other plant seeds) prior to sowing the seeds on media in a petri dish.  This is to reduce the possibility of contamination and bacterial growth that can present over the course of an experiment.  Although the quality of the seeds plays a large role in susceptibility to contamination, sterilization is still best practice to minimize risk.  

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.  70% ethanol is flammable and should be handled and stored appropriately. 

3.0: Definitions and Acronyms

  • ASOP: Astrobotany Standard Operating Procedure
  • EtOH: Shorthand for Ethanol
  • A. thaliana: Shorthand for Arabidopsis thaliana

4.0: Materials

Equivalent Materials may be used. 

  • 70% Ethanol
  • 1.5 mL Microcentrifuge Tubes
  • L-1000 Automatic Pipette or a disposable plastic transfer pipette
  • Filter Paper

5.0: Procedure

  1. Acquire the required amount of A. thaliana seeds and use a small piece of filter paper to funnel the seeds into a 1.5 mL microcentrifuge tube.
  2. Use an automatic or disposable transfer pipette to deliver enough 70% EtOH to fully cover the seeds in the microcentrifuge tube.
  3. Cap the tube and let the seeds sit in the 70% EtOH for approximately 2 minutes.  Feel free to gently shake or invert the tube.  Let the seeds settle to the bottom of the tube.
  4. Use the pipette to remove as much 70% EtOH as possible from the tubes, taking care not to remove any seeds. 
  5. Add a second wash of 70% EtOH to the seeds.
  6. Remove both the seeds and 70% EtOH from the 1.5 mL microcentrifuge tube using the pipette.
  7. Deliver the seeds and 70% EtOH onto a piece of sterile filter paper.  The seeds are now ready to be used or added to agar in a petri dish. 

Version History

  • Version 1 | Description: ASOP-0001 page created. | Effective: 16Jun2023
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