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Offlinenn-IlliniSpiralDMT
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Registered: 07/19/12
Posts: 380
Last seen: 7 years, 3 months
Trying to save my isolate PLEASE HELP!!!
    #18895424 - 09/26/13 06:49 PM (10 years, 4 months ago)

Okay so awhile back I made some agar plates with some random spores I had found of some very good lions mane I had grown a VERY VERY VERY long time ago, I am talking years here.

So I made 5 plates and chose the two that looked promising too me, I did a couple transfer and ended up with a nice plate that fruited within 10days, then I transfered that and fruited that, took the biggest fruit. Which was a nice 2 incher :laugh: and took an inside tissue of that fruit! No I had a good plate and it sectored beautifully, so I had a good plate wand was going to take different strands of mycelium from that plate and start a couple master jars to pick my favorite candidate with my desired  traits that I find ideal.

This is where the problem came, I had run out of dishes and ordered some new ones but they took forever, so I did something on Amaz**n and purchased these plates from a PRIME vendor, when I got the dishes I knew I had bad fate coming, they 60mm X 15mm which were way too small, but I used them anyway...

Went to the GloveBox to do work, I cleaned the GB with iso and let air dry, wiping my tools and equipment before I placed them in the glovebox and then wiping them again when I closed the GB. I did 10 plates of 10 different sectors, and then I closed them up, and left. I parafilmed them after I got done left them consolidate for 5 days and when I looked( I was expecting to see massive plates) they had some sort of bacteria or yeast on them... I was distraught.
I thought it was maybe because I left the lid open of my agar jar the entire time while I was doing my Agar to Agar transfer, but then I looked and saw that my entire jar of Agar had a gross smell to it and had shit growing on the top, but I did sterilize it for 40min at 15psi? SO I wonder if the contams came because of my poor procedure

So I was wondering If


a.) I could pour new dishes and pick up the pieces I placed on the contamed plates and transfer them to new ones. The pieces look fine as they have mycelium growing aerially,

b.) Just cut out small rice grain pieces out of the original agar ( it looks contam free but it does have a lot of holes in it from the pieces I took out) But still enough for 10 dishes.

or

C.) IM FUCKED



ANY HELP WOULD BE AWESOME THANKS GUYS

IS


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OfflineAmanita virosa
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Registered: 12/04/11
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Re: Trying to save my isolate PLEASE HELP!!! [Re: nn-IlliniSpiralDMT]
    #18895719 - 09/26/13 08:09 PM (10 years, 4 months ago)

Go back to the original plate. Make two plates from it. Pick the cleanest of the two and slant it. Then use it to run a sleeve of plates. Honestly  I doubt you will ever use a sleeve of plates unless you are a large commercial grower.  One plate equals six g1s equals 90 g2s. That's. shitload of spawn per plate pal.


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OnlineStromriderM
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Registered: 06/02/13
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Re: Trying to save my isolate PLEASE HELP!!! [Re: Amanita virosa]
    #18896118 - 09/26/13 09:31 PM (10 years, 4 months ago)

Hey Amanita I'm loving the dog food agar I got from you. It's working great! :thumbup:


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Offlinenn-IlliniSpiralDMT
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Registered: 07/19/12
Posts: 380
Last seen: 7 years, 3 months
Re: Trying to save my isolate PLEASE HELP!!! [Re: Amanita virosa]
    #18896318 - 09/26/13 10:12 PM (10 years, 4 months ago)

Quote:

Amanita virosa said:
Go back to the original plate. Make two plates from it. Pick the cleanest of the two and slant it. Then use it to run a sleeve of plates. Honestly  I doubt you will ever use a sleeve of plates unless you are a large commercial grower.  One plate equals six g1s equals 90 g2s. That's. shitload of spawn per plate pal.




So do you think that the fruit I took already is an isolate of its own after all those transfers since it was from a fruit I would assume so right, I figure it was growing in different sectors still? Thats what I was thinking too? And dog food agar? sounds interesting? Any old dog food? I've been using cornmeal, potato broth, and agar with a little karo in it for good measure. How are your dog food agar plate doing? And does that lead to darker agar?


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OfflineAmanita virosa
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Registered: 12/04/11
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Re: Trying to save my isolate PLEASE HELP!!! [Re: nn-IlliniSpiralDMT]
    #18897983 - 09/27/13 10:04 AM (10 years, 4 months ago)

yea, i would be surprised to see more than a few sectors from a clone of a fruiting body.  either way, its your only choice.  your contaminated plates are toast.  trash them.  just make some more plates from the clean origonal.  if they sector like crazy sub them out some more.  dog food agar is half ALL NATURAL (NO PRESERVATIVES) dry dog food and half agar by weight.  pulverize the dog food.  some good brands are Lassie brand and NUtri balance.  Works like a charm..


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Offlinenn-IlliniSpiralDMT
Maniac


Registered: 07/19/12
Posts: 380
Last seen: 7 years, 3 months
Re: Trying to save my isolate PLEASE HELP!!! [Re: Amanita virosa]
    #18898303 - 09/27/13 11:38 AM (10 years, 4 months ago)

Quote:

Amanita virosa said:
yea, i would be surprised to see more than a few sectors from a clone of a fruiting body.  either way, its your only choice.  your contaminated plates are toast.  trash them.  just make some more plates from the clean origonal.  if they sector like crazy sub them out some more.  dog food agar is half ALL NATURAL (NO PRESERVATIVES) dry dog food and half agar by weight.  pulverize the dog food.  some good brands are Lassie brand and NUtri balance.  Works like a charm..




Alright thanks buddy! SO any dog food that has no preservatives is viable for mycology? Im sure it will work great, and alright Ill toss my old plates. Really disappointing. Also what do you do in a glove box when you pour agar? DO you leave your agar jar container opened the entire time? Or do you open, pour a dish, close lid on dish, and then close agar jar, then repeat until your done with a sleeve. I think Ill take two samples and isolate like you previously stated.


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OfflineAmanita virosa
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Registered: 12/04/11
Posts: 2,458
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Last seen: 6 months, 17 days
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Re: Trying to save my isolate PLEASE HELP!!! [Re: nn-IlliniSpiralDMT]
    #18898370 - 09/27/13 11:57 AM (10 years, 4 months ago)

most important in a glove box is to glove and tyvek sleeve your hands.  yes you can leave the jar open when you pour the plates.  stack the plates 4 high in piles and grab the lid of the bottom plate with your hand over the top three empty ones.  pour in agar and release lid, they grab the lid of the third plate with the top two still in your hand, pour and drop lid.. then grab the second lid with the first plate still in your hand, drop the lid then grab the first lid and pour.  use this system for all the stacks.  if you have big hands you may be able to stack five but it is more awkward, esp in a glove box.  good luck


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Offlinenn-IlliniSpiralDMT
Maniac


Registered: 07/19/12
Posts: 380
Last seen: 7 years, 3 months
Re: Trying to save my isolate PLEASE HELP!!! [Re: Amanita virosa]
    #18898410 - 09/27/13 12:08 PM (10 years, 4 months ago)

Quote:

Amanita virosa said:
most important in a glove box is to glove and tyvek sleeve your hands.  yes you can leave the jar open when you pour the plates.  stack the plates 4 high in piles and grab the lid of the bottom plate with your hand over the top three empty ones.  pour in agar and release lid, they grab the lid of the third plate with the top two still in your hand, pour and drop lid.. then grab the second lid with the first plate still in your hand, drop the lid then grab the first lid and pour.  use this system for all the stacks.  if you have big hands you may be able to stack five but it is more awkward, esp in a glove box.  good luck




I have tyvek over my port holes and then I slip my gloves through. I used to attach my gloves but I heard that doesn't work as good. Thanks for all your help buddy


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