Griffitts:Seed scarification and germination

Home     Research     Lab Members     Protocols     Publications     Internal     Emeritus    

Alfalfa

Scarification

• Alfalfa seeds don't need to be scarified.

Germination

• Calculate the number of plants you need
• Weigh out ten times more than the number you need
  • Use GT13R Plus
  • 1.7 g is approximately 1000 seeds
• Place in a smooth sterile flask
• Add 70% ethanol to substantially cover the seeds
• Place on a shaker at room temperature at 150 rpm for 20 minutes
• Decant the ethanol and wash the seeds once with sterile dH₂0
• Add Clorox bleach to substantially cover the seeds
• Place on a shaker at room temperature at 150 rpm for 20 minutes
• Decant the bleach and wash the seeds 4 to 5 times with sterile dH₂0
• Add sterile dH₂0 to substantially cover the seeds
• Place on a shaker at room temperature at 150 rpm for at least 4 hours (preferably 8)
  • It's a good idea to wash the seeds with sterile dH₂0 once during this time
• Decant the sterile dH₂0 and add fresh sterile dH₂0
• Dump into 100×25 mm Petri dishes
  • Seeds should be in a monolayer with a small amount of space between them
• Pull off excess water with a pipet
  • Tip the plate and give it a few seconds to be sure to get all the excess water
• Wrap plate(s) in Parafilm
• Store upside-down in a drawer for 24 hours