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-   -   Very Easy Sterilisation Method - My Seedlings (https://www.orchidboard.com/community/propagation/43343-easy-sterilisation-method-seedlings.html)

Phalguy 02-05-2013 07:12 PM

Quote:

Originally Posted by Call_Me_Bob (Post 550166)
Nice!!!! How did you sterilize when you replated?


I used the same technique, bleach fume ! On 4 flask, only one got contaminated and 2 dried up over time. It's the only flask surviving so far....

It,s a very long process but soooooooo fun to do !

Call_Me_Bob 02-05-2013 11:51 PM

Quote:

Originally Posted by Phalguy (Post 550339)
I used the same technique, bleach fume ! On 4 flask, only one got contaminated and 2 dried up over time. It's the only flask surviving so far....

It,s a very long process but soooooooo fun to do !

So you were able to replate with this method as well? Awesome I will have to give it another go! I thought to replate you would still need a glove box or something similar.

Phalguy 02-06-2013 05:25 PM

I find that using a glove box with this technique minimize the contamination... but you don't have to... I have use the water steam ( on a wok ) instead of glove box with good results..

Call_Me_Bob 02-25-2013 01:42 PM

DO you guys think this would work for doing a stem prop? Any thoughts on that?

biofuel 04-09-2013 07:48 AM

Today I try gaseous sterilization.

I used seeds Epipactis helleborine collected in November 2012.
Pad was soaked in 5% NaOCl – 3 hours.
Seeds were sown on modified BM-2 (with 25% coconut water and 10 grams - not 20 - sucrose per liter).
All jars have a lid with a cotton plug.
Medium sterilized for one hour in 180C, electric oven.
I made 6 jars and now I'm waiting for contaminations ;)
Pictures

camille1585 04-09-2013 07:56 AM

Your photos are not appearing. Unfortunately you are not allowed to post photos or links until you have made 5 posts. Post a few more times, and then it's good!

biofuel 04-09-2013 08:04 AM

It's explain why I have a small problems with sharing the photos, thank you ;)

biofuel 04-19-2013 03:02 AM

After ten days I have 8 jars with bacterial contamination...
In two jars I dont see any signs of contamination. I don't know what I did wrong.
I have a suspicion:
Probably under the orchid seeds could be a bacterial spores, and Cl2 can't kill them, because a part of the surface seed coat didn't avaliable to sterilization agent.

escualida 04-19-2013 03:20 AM

Quote:

Originally Posted by biofuel (Post 568127)
After ten days I have 8 jars with bacterial contamination...
In two jars I dont see any signs of contamination. I don't know what I did wrong.
I have a suspicion:
Probably under the orchid seeds could be a bacterial spores, and Cl2 can't kill them, because a part of the surface seed coat didn't avaliable to sterilization agent.

That makes sense, I wonder if there's a way of getting under there...

biofuel 04-19-2013 03:52 AM

Epipactis helleborine have a bigger seeds than other orchids, so I think it's a cause.
Maybe first sterilization seeds in 70% ethanol could solve this problem.
I must try it in this weekend.


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