Griffitts:Seed scarification and germination: Difference between revisions
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* Remove the sulfuric acid | * Remove the sulfuric acid | ||
** Discard it into the sulfuric acid waste container in the fume hood | ** Discard it into the sulfuric acid waste container in the fume hood | ||
* Immediately rinse the seeds | * Immediately rinse the seeds five times with dH<sub>2</sub>0 | ||
* Add 5 mL of bleach | * Add 5 mL of bleach | ||
* Place them back onto the shaker for 3 minutes | * Place them back onto the shaker for 3 minutes | ||
* Rinse | * Rinse 6–8 times with dH<sub>2</sub>0 | ||
* Pour the seeds and some dH<sub>2</sub>0 into small plates | * Pour the seeds and some dH<sub>2</sub>0 into small plates | ||
* Parafilm and store in the fridge 24 hours to allow imbibement | * Parafilm and store in the fridge 24 hours to allow imbibement |
Revision as of 12:30, 4 April 2008
Alfalfa
Note: Alfalfa seeds don't need to be scarified
- 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
- 70% ethanol is next to the scales
- 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
Other Medics
- Place twice as many seeds as you wish to plant into a 50 mL Erlenmeyer flask that has been covered with foil and autoclaved.
- To each of the flasks add 5 mL of sulfuric acid (H2SO4, in the fume hood—be careful!)
- Place on the shaker for the appropriate time (see table below)
- Remove the sulfuric acid
- Discard it into the sulfuric acid waste container in the fume hood
- Immediately rinse the seeds five times with dH20
- Add 5 mL of bleach
- Place them back onto the shaker for 3 minutes
- Rinse 6–8 times with dH20
- Pour the seeds and some dH20 into small plates
- Parafilm and store in the fridge 24 hours to allow imbibement
- Be sure not to have excessive amounts of water in the plate
- Remove the seeds from the fridge and remove the water
- Parafilm the mini-plates, invert and place at room temperature in the dark for one day
Note: some plants may take a little longer to germinate
species | time | notes |
---|---|---|
arabica | 15 minutes | |
constricta | 10 minutes | |
disciformis | 10 minutes | |
heyniana | 20 minutes | |
italica | 10 minutes | |
lesinsii | 10 minutes | |
lupulina | 20 minutes | |
polymorpha | 20 minutes | |
praecox | 8 minutes | refrigerate in 15 mL dH2O for 36–48 hours |
rugosa | 30 minutes | |
tenoreana | 10 minutes | |
truncatula A17 | 10 minutes |
Lotus japonicus
- Place twice as many seeds as you wish to plant into a 50 mL Erlenmeyer flask that has been covered with foil and autoclaved.
- To each of the flasks add 5 mL of sulfuric acid (H2SO4, in the fume hood—be careful!)
- Place on the shaker for 20 minutes
- Remove the sulfuric acid
- Discard it into the sulfuric acid waste container in the fume hood
- Immediately rinse the seeds once with dH20
- Add 5 mL of bleach
- Place them back onto the shaker for 1 minute
- Rinse five times with dH20
- Pour the seeds and some dH20 into small plates
- Parafilm and store in the fridge 24 hours to allow imbibement
- Be sure not to have excessive amounts of water in the plate
- Remove the seeds from the fridge and remove the water
- Parafilm the mini-plates, invert and place at room temperature in the dark for one day