The following equipment and consumables are readily available near a kitchen range:
- cooking pot
- Alcohol burner
- Small screw jars
- Test tubes with finished culture medium
- Vaccination loop
- Sharp knife or scalpel
- coffee filter
- Cotton pads
- Distilled water
- Ethanol (70 percent)
- Hydrogen peroxide (3 percent)
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Hydrogen peroxide is used for disinfection and is available in a ready-to-use 3 percent dilution in pharmacies and online shops. You can read here how easy it is to prepare nutrient media.
Obtaining and preparing seeds from a seed pod - How to do it right
If manual pollination was successful, wait for a seed pod to pop open. Cut these off, shake the seeds on coffee filter paper and shape them into an envelope, closed with pins. The seeds must be sterilized before sowing on the culture medium. That is how it goes:
- Pour the water into the saucepan up to 3 cm high and bring to the boil
- Clean the wire rack with ethanol and place on the saucepan
- Fill a screw jar about 1 cm high with hydrogen peroxide
Using the tweezers, place a seed envelope in the solution for 10 minutes. Repeated swirling ensures that all seeds are moistened.
Apply seeds to the medium
The following work is all carried out over a constant stream of steam that simulates the conditions of a sterile workbench. Follow these steps:
- Place a cotton swab soaked in ethanol and a glass of distilled water on the wire shelf
- Remove the seed envelope from the glass with the tweezers in a stream of steam and place it in the distilled water
- Swirl the seed envelope in the water and place it on the cotton pad to open it with the tweezers and scalpel
- Take a test tube with a nutrient medium, open it in a stream of steam and place it on a cloth soaked in ethanol
With the help of the inoculation loop, you can apply the seeds from the envelope directly to the nutrient medium and distribute them there. Then close the test tube again with the stopper and put an aluminum foil cap on, ideally closed with a rubber ring. It is important to note that before and after each work step you clean the tool with ethanol or briefly flame it over the alcohol burner.
After sowing, place the culture vessels at a warm 25 degrees Celsius in a bright location without direct sunlight. Here there should not be any strong temperature fluctuations that could trigger a pressure change in the test tubes.