Griffitts:Seed scarification and germination: Difference between revisions
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==Other Medics== | ==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. | * 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 H<sub>2</sub>SO<sub>4</sub> | * To each of the flasks add 5 mL of sulfuric acid (H<sub>2</sub>SO<sub>4</sub>, in the fume hood—be careful!) | ||
* Place on the shaker for the appropriate time (see table below) | * Place on the shaker for the appropriate time (see table below) | ||
* Remove the sulfuric acid | * Remove the sulfuric acid | ||
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* Immediately rinse the seeds once with dH<sub>2</sub>0 | * Immediately rinse the seeds once with dH<sub>2</sub>0 | ||
* Add 5 mL of bleach | * Add 5 mL of bleach | ||
* Place them back onto the shaker for | * Place them back onto the shaker for 2-3 minutes | ||
* Rinse five times with dH<sub>2</sub>0 | * Rinse five 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 | ||
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| 10 minutes | | 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 (H<sub>2</sub>SO<sub>4</sub>, 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 dH<sub>2</sub>0 | |||
* Add 5 mL of bleach | |||
* Place them back onto the shaker for 1 minute | |||
* Rinse five times with dH<sub>2</sub>0 | |||
* Pour the seeds and some dH<sub>2</sub>0 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<br> |
Revision as of 11:34, 13 February 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 once with dH20
- Add 5 mL of bleach
- Place them back onto the shaker for 2-3 minutes
- 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
Note: some plants may take a little longer to germinate
species | time |
---|---|
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 | 10 minutes |
rugosa | 30 minutes |
tenoreana | 10 minutes |
truncatula A7 | 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