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flipsidetrue




Registered: 05/26/13
Posts: 364
Loc: Earth
Last seen: 7 years, 10 months
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That's what I'm saying! I'm over here growing grey hairs about to look like Albert Einstein over here
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MUSH HEAD420
Kush Commander



Registered: 09/14/08
Posts: 921
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Well if it happened with the spores and the sample so it did not come from the syringe or the sample.. unless one introduced it to the other.
I am guessing it is in the environment, try and do it outside or something and then seal your agar plates with mircopore tape.
Edited by MUSH HEAD420 (10/31/13 11:29 PM)
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cronicr



Registered: 08/07/11
Posts: 61,436
Loc: Van Isle
Last seen: 2 years, 8 days
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Do u have more fruits/tissue samples? Did you take prints?
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  It doesn't matter what i think of you...all that matters is clean spawn I'm tired do me a favor
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flipsidetrue




Registered: 05/26/13
Posts: 364
Loc: Earth
Last seen: 7 years, 10 months
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Re: hairy mycelium? [Re: cronicr]
#19067054 - 10/31/13 11:30 PM (10 years, 2 months ago) |
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yes i have a couple more prints and I'm down to 1 more fruit and I'm not to confident about the prints being clean
Edited by flipsidetrue (10/31/13 11:31 PM)
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cronicr



Registered: 08/07/11
Posts: 61,436
Loc: Van Isle
Last seen: 2 years, 8 days
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Sometimes prints can be easier to clean with wild fruits but with tissue i use to use a method rr wrote with a series of rinses in sterile water in half pints where u just sterilize a few jars of water drop your fruit in and shake, u can add laundry bleach to it if you want but I never did, if I still had problems I turn to cardboard to get some growth and clean that on agar, just my two cents but just be more persistent then the contam lol
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  It doesn't matter what i think of you...all that matters is clean spawn I'm tired do me a favor
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flipsidetrue




Registered: 05/26/13
Posts: 364
Loc: Earth
Last seen: 7 years, 10 months
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Re: hairy mycelium? [Re: cronicr]
#19067230 - 11/01/13 12:07 AM (10 years, 2 months ago) |
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hmmmmm interesting... i think i might try to mix up some h202 solution to help fight the contaminants and then keep transferring until i get a solid healthy growth.. what kind of h202 mixture would you recommend?
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MUSH HEAD420
Kush Commander



Registered: 09/14/08
Posts: 921
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1:10 - 2:10 of 3% peroxide, you can use 1/2 strength 3% peroxide at first to kill the stuff. do not over or under do it.
Edited by MUSH HEAD420 (11/01/13 12:17 AM)
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bodhisatta 
Smurf real estate agent



Registered: 04/30/13
Posts: 61,889
Loc: Milky way
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I've been told over and over by the likes of RR and a few trusted cultivators to not shock the mycellium with H2O2 when using clone tissue to agar. You want the healthy myc to out run the contams. Cobweb is tricky but if you're seeing it with spores and your clone tissue I have a feeling it's your sterile technique. It's probably getting in there from some part of pouring your plates or when you open them to put the clone on. Double check your SAB/sterile techniques.
If you want to use the clone tissue from that outdoor fruit. Rip the fruit in half inside your SAB from the base. You should have virgin never seen air tissue on the inside of the stem that you can carefully transfer to the agar. You want to be extra careful not to let any part of your tool touch the outside surface of the fruit just get inside the stem tissue.
Unfortunately I dont think h2o2 is going to help you. It never helped me out and actually made the cobweb run the dishes even more vigorously since it's pretty hard to catch every last drop of cobweb with an h2o2 dip.
You're going to want to make as many dishes as you can from the clone tissue or spore print to start. That way if you make 6 and only 1 is success you can transfer that one to 6 more and toss the 5 that sucked. When starting from such dirty original samples you're going to want probability and numbers on your side so do 10 dishes if you can.
Use parafilm or saran wrap to wrap the dishes.
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SpitballJedi
Ancient Astronaut



Registered: 10/13/12
Posts: 8,598
Loc: Nibiru
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Have you tried this:
Go through the whole agar process as normal, except don't actually inoculate.
Do the open and close the dish, parafilm, the whole nine. Let the dish sit and see if it contaminates.
-------------------- The Basics A little civility goes a long way The Noob Forum The Hammock Hangers' Forum
Edited by SpitballJedi (11/01/13 08:55 AM)
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flipsidetrue




Registered: 05/26/13
Posts: 364
Loc: Earth
Last seen: 7 years, 10 months
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@spitballjedi: I've done something very similar to that actually, when i first got my MEA i went ahead and poured into several dishes so all i would have to do was inoculate or place tissue into, and the ones that i poured and left alone actually had no contaminants
this is the SAB i'm using
Edited by flipsidetrue (11/01/13 12:11 PM)
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flipsidetrue




Registered: 05/26/13
Posts: 364
Loc: Earth
Last seen: 7 years, 10 months
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Btw let me add i live in a household with 4 dogs 2 cats that are constantly running around
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Yuri.Pono
MAD SCIENTIST


Registered: 10/28/13
Posts: 279
Loc: SO HIGH O
Last seen: 9 years, 9 months
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Quote:
flipsidetrue said: i really didn't want to order more spores because to do so consists of me having to get a p.o box in another state and picking them up from there 
that does suck
-------------------- Links To Success finger print: A9E348E5 2A22AFB6 F3676613 A34B1454 7C36ABC8 want to chat? got xxmp/pidgin/otr PM me for info
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SpitballJedi
Ancient Astronaut



Registered: 10/13/12
Posts: 8,598
Loc: Nibiru
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Just for the sake of being thorough, When you poured your plates before and just let them sit, were they wrapped in the same tape as the above pictures? Were they stored/kept exactly the same way as your inoculated dishes.
I'm just trying to help see if there is some subtle detail that's different.
In my experience, most people don't have the exact same contams when using spores AND tissue unless there is something wrong about the technique. Sometimes it's very subtle.
-------------------- The Basics A little civility goes a long way The Noob Forum The Hammock Hangers' Forum
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flipsidetrue




Registered: 05/26/13
Posts: 364
Loc: Earth
Last seen: 7 years, 10 months
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the plates that i did pour and let sit i didn't wrap them in any tape or Saran wrap because i read somewhere that it wasn't necessary if tissue/ spores weren't added to them, i actually just sat them in my closet and they had no signs of contams until about 48 hours after i added the tissue/ spores
Edited by flipsidetrue (11/01/13 01:46 PM)
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SpitballJedi
Ancient Astronaut



Registered: 10/13/12
Posts: 8,598
Loc: Nibiru
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Did you leave the poured/blank ones in the original bag or something?
-------------------- The Basics A little civility goes a long way The Noob Forum The Hammock Hangers' Forum
Edited by SpitballJedi (11/01/13 02:25 PM)
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flipsidetrue




Registered: 05/26/13
Posts: 364
Loc: Earth
Last seen: 7 years, 10 months
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nope i didnt even do that much i just sat them on a shelf in my closet
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SpitballJedi
Ancient Astronaut



Registered: 10/13/12
Posts: 8,598
Loc: Nibiru
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-------------------- The Basics A little civility goes a long way The Noob Forum The Hammock Hangers' Forum
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flipsidetrue




Registered: 05/26/13
Posts: 364
Loc: Earth
Last seen: 7 years, 10 months
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i know right? so i think what I'm going to do is spray the inside with some bleach/water solution,then go to town with my brand new x ac-to knife after flame sterilization of course. But before i do anything i have a concern about something which is i had a brand new sleeve of Pre-sterilized petri dishes that i opened inside the SAB but i didnt use all 20 of them, I'm thinking i might have messed up by doing so?
Edited by flipsidetrue (11/01/13 05:42 PM)
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MUSH HEAD420
Kush Commander



Registered: 09/14/08
Posts: 921
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use h2o2 not bleach.. imo
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flipsidetrue




Registered: 05/26/13
Posts: 364
Loc: Earth
Last seen: 7 years, 10 months
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alright note taken i wanna thank everyone that chimed in on this post for your advice and help! MUCH LOVE!
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