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InvisibleTFI
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How long should i wait before throwing away agar plates
    #23191523 - 05/05/16 08:28 AM (8 years, 8 months ago)

Ok heres what happened. after i had a successful grow months and months ago i got lazy and let the next set of spawn jars sit for 2-3 weeks not caring, well I finally spawned them only to see bacteria and trich take its tole. My last tub pinned amazingly but thats all it did. Pin set was starting to turn blueish blackish crap.. Soooo i mixed up some agar plates and took transfers from said tub and a spawn jar that i held aside... I took three pins, split them in half and took a cut from the inside. One transfer from wbs spawn jar to agar, its been four days and the only thing that i see is the birdseed fluffing back up but no growth towards the agar itself. The other samples turned blue from the heat of the scalpel and have yet to show anything at all... Did i kill the mycelium? How the fuck do you flame sterilize and then cool down said tool without contaminating it? Should i just keep waiting? How long till I should dump them and restart? I have done agar in the past with good success but cannot remember the colonization times

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InvisiblePastywhyteMDiscord
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Re: How long should i wait before throwing away agar plates [Re: TFI]
    #23191534 - 05/05/16 08:36 AM (8 years, 8 months ago)

Cool down the scalpel by cutting into the receiving plate. Then take the biopsy.

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InvisibleTFI
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Re: How long should i wait before throwing away agar plates [Re: Pastywhyte]
    #23191625 - 05/05/16 09:27 AM (8 years, 8 months ago)

Ok so just so im clear here, flame sterilize, get in sab, open receiver jar, dip scalpel, take the cut from said tissue then re dip leaving said sample in agar? Or could i make a blank plate to do my cooling?

Im thinking now I just have a bunch of dead tissue lmao.. Also i am using your tek so thank you very much for that handy info

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OfflinePussyFart
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Re: How long should i wait before throwing away agar plates [Re: TFI]
    #23191682 - 05/05/16 09:54 AM (8 years, 8 months ago)

I buy blades off ebay from china by the thousand for super cheap.

They come individually wrapped and sterilized.

I use a new blade for every plate.

No flaming. So much easier lol.

Edited by PussyFart (05/05/16 09:54 AM)

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OfflinePussyFart
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Re: How long should i wait before throwing away agar plates [Re: PussyFart]
    #23191725 - 05/05/16 10:09 AM (8 years, 8 months ago)

Slice the same plate you are using for the transfer.

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InvisibleTFI
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Re: How long should i wait before throwing away agar plates [Re: PussyFart]
    #23191734 - 05/05/16 10:12 AM (8 years, 8 months ago)

Thats a good idea! And ok ill try that today. Just kinda scares me to open up receving plate twice lol damm contams.. I gotta invest in a laminar flow hood

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OfflinePussyFart
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Re: How long should i wait before throwing away agar plates [Re: TFI]
    #23191799 - 05/05/16 10:34 AM (8 years, 8 months ago)

If you fail this in a SAB you will fail with a hood.

With good sterile technique, you are golden.

You can stick me in a room filled with trich and with my SAB I can do 100 transfers and still come out a winner.

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InvisibleSupalemonhaze
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Re: How long should i wait before throwing away agar plates [Re: PussyFart]
    #23192487 - 05/05/16 02:18 PM (8 years, 8 months ago)

I feel you man, I never had any luck in a SAB.

Sometimes mycelium takes a while to recover if the transfered mycelium is very small. It's not uncommon for it to start recovering 5+ days later. IME clones turn blue even when the scalpel is cool, it's just a response to being damaged.

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InvisibleNeomorph
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Re: How long should i wait before throwing away agar plates [Re: Supalemonhaze]
    #23192794 - 05/05/16 04:03 PM (8 years, 8 months ago)

If one culture will colonize slow on agar does that mean that it will colonize slow even on grains ?

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InvisibleSupalemonhaze
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Re: How long should i wait before throwing away agar plates [Re: Neomorph]
    #23194476 - 05/06/16 04:02 AM (8 years, 8 months ago)

If it is a multistrain culture chances are there is something wrong with it's medium. Contamination, too much nutes or the agar being too soft comes to mind. If you have been using the same recipe for some time it could also have gotten used to it too much. You should have at least 2 recipes to keep the cultures vigorous but having more always helps.

Cultures might do better on some recipes than on others as well, so you might want to experiment with your ingredients a bit.

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InvisibleTFI
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Re: How long should i wait before throwing away agar plates [Re: Supalemonhaze]
    #23194734 - 05/06/16 07:51 AM (8 years, 8 months ago)

I feel as if im sterile, but i believe a flow hood would only increase the potential success rate. if i drop something or move around too much I usually take that jar or plate and set it aside to see if i messed it up. 9/10 are usually fast recovering great spawn. I believe that i just let the previous jars sit to the point of primordia in the jars, I broke em up an soawned any way which I believe led me here.

Supalemonhaze i was gonna say the same thing because its day 5 and their starting to show some white recovery lol,

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InvisibleSupalemonhaze
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Re: How long should i wait before throwing away agar plates [Re: TFI]
    #23194880 - 05/06/16 09:06 AM (8 years, 8 months ago)

For the most part what PF said is accurate, you will still fuck up with a hood. Only time I would say a hood is what you need is if you have no success whatsoever with a SAB.

Before I built mine I tried everything, I even worked with plates upside down. What the reason was is still a mystery to me, but months later when I finally built my hood I went through the few uncontamined agar plates I had as well as some unused no-pours and I found out I had mites in every single petri so it's possible that it was a problem from day one. I still believe that it couldn't have been the only problem, mites take some time before they make it inside a plate. Thankfully, I haven't used a SAB since then and I have no intention of finding out what the problem was.

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