The Orchid Forum, for the UK and Europe (previously known as The UK Orchid Forum) • View topic - How to Prevent Steam-Cooking Disaster?
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PostPosted: Fri Feb 14, 2014 2:09 pm 
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Hi,

I use small glass containers for the flasking. I create the mixture, and then steam-cook the flasks for 45 minutes to ensure any fungi spore or any bacteria is death.

But my problem is, that after the steam-cooking the germination mix is not just at the bottom of the glass, but even the walls are covered by many smaller and bigger pieces of the mixture, blocking free view and light.

How to prevent this?

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PostPosted: Mon Feb 17, 2014 12:37 am 
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I think it is best to use jars with plastic lids so you can sterilize in a microvave, it only takes about 3 minutes!

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PostPosted: Mon Feb 17, 2014 8:05 am 
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Microwave? I didn't knew this way yet! Thanks. I will consider it. 3 minutes on 100%? And can it really kill all fungi spores and all bacterias?

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PostPosted: Mon Feb 17, 2014 9:20 pm 
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You must have the lids not fully tight to allow for expansion, You put the microwave on until the agar starts to boil/bubble, about 3-4 minutes, turn off, after this time allow a few minutes to cool slightly before tightening the lids. This method I am sure is described with the flasking protocol you downloaded from The orchid & gardening forum on facebook.

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PostPosted: Wed Feb 26, 2014 11:01 pm 
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Thanks a lot! At the moment, all my flasks come with metal covers.

If I find a source for long-term plastic-cup glass containers, I will for sure give a try to the microwave. Where do you get your germination containers from? Can you give an URL? I might want to order from them, too.

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PostPosted: Fri Aug 01, 2014 11:53 pm 
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Your, problem with vitro forming up the inside of your mother flask. Try wedging a rag between the flasks or use a string around the neck of the flask. If the flask rattle or sway inside the autoclave/pressure cooker this can cause the vitro to run up the sides.

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PostPosted: Wed Mar 30, 2016 8:41 pm 
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Good advise! Thank you.

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PostPosted: Wed Mar 30, 2016 10:30 pm 
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If you are using a pressure cooker as your auto clave, and vent the pressure off to quickly this can happen.

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PostPosted: Sun Apr 03, 2016 7:14 pm 
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I use the pressure cooker method having jars with metal lids, I steam them for 15 minutes once to temperature/hissing, that time is sufficient for 100% success sterilisation of the jars for me.

I then let cool over 5-10 minutes before opening to avoid excessive bubbling of the jars, very little spillage on the sides of the jars.


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PostPosted: Sun Apr 03, 2016 8:40 pm 
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I also use a pressure cooker at 12psi for 20 min once the pressure is achieved with 100% success.
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