If you want to flask dry seeds you have to wait till your capsule opens. When the capsule starts to dehisce cut the capsule and shake the dry seeds out on a sheet of paper. Before you start sterilizing your seeds you should remove all contaminations (parts of the capsule, pollen tubes, ...)
necessary Tools:
necessary articles of consumption:
You can find the sources of supply at Equipment.
|
Advantages of using dry seeds:
Disadvantages of using dry seeds:
|
Preparing the flasking area:
We us the steam above a pot with boilding water to provide sterile conditions. To minimize the risk of contaminations you should reduce draft in your room as much as possible. Close all windows and doors while you are flasking. In the picture below you can see our preferred arrangement of tools (for right-handed person).
Making a 2% hydrogen peroxide desinfection solution:
To reduce the concentration of the bought hydrogen peroxide (e.g. 35% H2O2) to the required 2% we have to add some distilled water to the high concentrated solution. With the formula below you can calculate how much distilled water you have to add to the high concentrated solution to get a 2% solution.
Wanted quantity of 2% hydrogen peroxide: |
50 ml |
Formula: quantity high% = (total quantity * %low) : %high = (50 * 2) : 35 = 2,86 ml |
|
Next steps:
Open the bottle with 70% ethanol and place you forceps or repaltingtool into it. Fill about 3 cm water in your pot and turn on your oven. The temperature of the boilding water must be high enough to produce a steady flow of steam. As soon as the water starts to boil, take a kitchen paper, soak it with 70% ethanol and use it to clean the grill. When you finished cleaning place the grill on the pod.
Sterilization of dry seeds:
Pack your seeds in a small filter paper envelope like the pictures below show.
Fill about 1 cm hydrogen peroxide in a screwable flask and put the filter paper envelope into the flask. Screw down the lid and agitate the flask for 10 minutes to make shure that the envelope has as much contact with the sterilization solution as possible.
Seed sowing:
The following steps must be done in the sterile area (steam). Open test tubes and their cotton plugs have to stay in the steam till the test tube is closed again.
After sterilizing the seeds you can start to transfer them into your test tubes. Put on your gloves, take the flask containing sterile distilled water and open it in the steam. Put down the flask on the grill.
Now take the flask where the seed envelope is swimming in and open it in the steam. Flame your forceps and transfer the envelope to the sterile distilled water.
After rinsing the envelope for some seconds in sterile distilled water move the envelope with a flamed forceps to the ethanol soaked kitchen paper (on the grill). Flame a scalpel and open the envelope.
Hold your scalpel and the forceps for a short moment in the boiling water and bring them back into 70% ethanol. Take a test tube and remove the aluminium foil cap. Place the cap close to the pod on a kitchen paper which is soaked with 70% ethanol.
Take the forceps out of the 70% ethanol and flame it. Remove the cotton plug with the flamed forceps and place the plug on the grill.
Flame your replating tool and pick up some seeds with it. Transfer the seeds directly into the test tube on the media. After bringing the seeds on the media hold the replating tool for a short moment in the boiling water and bring it back into 70% ethanol.
Pick up the forceps which is holding the cotton plug and put the plug back into the test tube. Hold the forceps for a short moment in the boiling water and bring it back into 70% ethanol.
Flame the cotton plug.
Put the aluminium foil cap on the test tube. To make shure that the foil cap does not move around we put a rubber band around it.
Some hints:
Further care:
We place our flask at our windowsill like the picture below shows. The temperature is about 20 °C. It is very important to prevent direct sun because the seeds in the flasks will become to hot if they get direct sun. If you have no windowsill available you can use a 20 Watt fluorescent tubes. We advice you to place the flasks on different places, so you can get a feeling which species needs which conditions to germinate.
