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Moderator: Matt

By Doomsday
Posts:  621
Joined:  Sun Mar 07, 2010 3:11 pm
#60533
Ok so even on my second attempt, I seem to have a difficult time doing all the sterilization. I used two bais stem pieces where the auxillary buds are supposed to be. It is on a BAP media now and everything is done, except these are my problems:

2. I hve 1 jar being used.
2. i have 3 other jars with nothing in them but they have gelled media
4. i have nearly a liter of liquid media without agar.

can the extra be saved> if yes then how?


Also, this is how i did my sterilization of everything

The tools:
dipped in bleach (my freakin exacto nife is now rusty so i hope rust wont kill the plants.... i rinsed it and bleached it agn and alohol and everything to kill bacteria on it)
and then sit in alcohol

i shook them off well before holding the explants for their final carry into the media jar.

the plants

i took the basi stem and cut off all of the leaves.

i then soaked it in 70% alcohol for 10 seconds, and then in 10% bleach for several minutes
i rinsed i in a jar of steril water for a couple minutes and then rinsed it again in another baby food jar of sterile water


i then used my alcohol dipped forceps and knife to cut of the ends that the bleach had made contact with, and split the explant in two (so i have 2 Y shaped explants) and placed them on the media

I wrapped the lid of the jar in saran wrap and put it in a ziplock bag

CShould this work??

This is the same procedure I did with my sundew and vft explants, and they have been on 1/2 strength media for about 1 week now. the vft leaf has a tiny bit of darkening at the end where it is cut open, but it doesnt look like its blackening. the sundew was an unculred leaf and around the bent part, the leaf is turning brown




SHould i give these a chance or just throw them away becaus they will die definately?

ALso, is this procedure good for the future, or wil i need to be far more exact and careful....


Also, al of this was done indoors in my ten gallon fish tank turned on its side and sprayed with 70% alcohol.
My jaws is very droopy....

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