Sterilizing the Surface of Seeds by Chlorine Gas
Lynn Doran
Abstract
This protocol sterilizes the surface of seeds for use in clean culture conditions without damaging the seeds germination or growth ability.
Steps
Move an aliquot of seeds to be sterilized to a 2 mL microcentrifuge tube. Label clearly.
Document the seed vials identifications and order via photograph or notetaking.
Place a desiccator in the chemical fume hood. Adjust the fume hood sash to a safe height.
Place the seed rack in a desiccator with no desiccant. Open the tube lids.
Fill a beaker with about 100mL of bleach, using graduations on the side of the beaker is sufficiently accurate for this purpose.
Place the beaker of bleach in the desiccator.
Quickly pipet 3mL of hydrochloric acid into the beaker of bleach.
Close the lid of the desiccator. If the desiccator contains any vacuum lines or vents, ensure that those valves are all in the closed position.
Allow the seeds to be exposed to the chlorine gas for a minimum of 3 hours.
Move the sealed desiccator to a biological safety cabinet that has been properly sterilized.
Open the desiccator slowly.
Close the seed vials. Seed vials can be stored outside the biological safety cabinet but should only be opened in properly sterilized biological safety cabinets or laminar flow hoods to ensure that they remain sterile.
