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OfflineTakeatrip
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Registered: 09/10/23
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BRF cake contamination cause with Golden Teacher?
    #28636525 - 01/27/24 07:33 AM (1 day, 27 minutes ago)



I am trying to eliminate what is happening whenever I try to get a certain spore to germinate. I get contamination each time I try whether it be in petri dishes or in BRF cakes.
The jars are prepared as per the guide on here and I leave them to cool overnight. I put them into the SAB the next day with gloves on and heat the needle outside of the SAB until red hot then quickly pass it through the hand holes and put to one side on a sterile glass dish. I then get my syringe ready by shaking it outside the SAB then passing it through the hand holes. I attach the needle then inject through the pre made holes in the lids and only about 2 drops or so. This has worked well for other strains I have without an issue but the Golden Teacher just keeps getting mould. I have tried ordering a new syringe of these spores from a totally different vendor so will be interesting next week to see what those results will be.
How do you guys get Golden Teacher to germinate successfully?
Weird thing is the syringe is loaded with spores as you can see them which is more promising than my last one.


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OfflineScrewup
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Registered: 01/27/22
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Re: BRF cake contamination cause with Golden Teacher? [Re: Takeatrip]
    #28636527 - 01/27/24 07:36 AM (1 day, 24 minutes ago)

Yeah that’s fucked.


Your golden teacher syringe probably just contaminated. Try a new vendor.


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Edited by Screwup (01/27/24 07:36 AM)


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InvisibleLadysKnight
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Registered: 10/09/15
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Re: BRF cake contamination cause with Golden Teacher? [Re: Takeatrip] * 1
    #28636529 - 01/27/24 07:40 AM (1 day, 20 minutes ago)

I could be misunderstanding, but it sounds like you flame the needle, let it cool, then attach it to the syringe?

I would attach the needle, shake then flame.

Either way, it looks like the syringe is fucked. I don't think it's you.


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OfflineBrokenHeart
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Re: BRF cake contamination cause with Golden Teacher? [Re: Takeatrip] * 2
    #28636533 - 01/27/24 07:42 AM (1 day, 17 minutes ago)

The GT syringes I bought for study barely had anything visible. Having many visible spores sounds like a good idea but with the larger spore load the greater the chance for contamination.  In your situation I would use the smallest amount I could on agar (probably 6-10 plates) and then transfer as soon as possible to try to separate good from bad.


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OnlineThe Tao
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Registered: 09/12/19
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Re: BRF cake contamination cause with Golden Teacher? [Re: Takeatrip]
    #28636538 - 01/27/24 07:48 AM (1 day, 12 minutes ago)

Why are you sterilizing the needle, putting it down then attaching the needle to the syringe?
Attach the needle to the syringe then heat the needle.


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InvisibleLadysKnight
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Re: BRF cake contamination cause with Golden Teacher? [Re: The Tao] * 2
    #28636558 - 01/27/24 08:03 AM (23 hours, 56 minutes ago)

Another thought, maybe just the pic, but that dry verm layer looks too thick. It needs to be thin enough to be able to poke the needle through to the brf. If you are injecting into the dry verm layer, this could be part of the problem.


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OfflineTakeatrip
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Registered: 09/10/23
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Re: BRF cake contamination cause with Golden Teacher? [Re: LadysKnight]
    #28637536 - 01/28/24 12:34 AM (7 hours, 25 minutes ago)

Quote:

LadysKnight said:
Another thought, maybe just the pic, but that dry verm layer looks too thick. It needs to be thin enough to be able to poke the needle through to the brf. If you are injecting into the dry verm layer, this could be part of the problem.



I have needles long enough to get below the verm layer so its ok


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OfflineTakeatrip
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Registered: 09/10/23
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Re: BRF cake contamination cause with Golden Teacher? [Re: The Tao]
    #28637540 - 01/28/24 12:39 AM (7 hours, 21 minutes ago)

Quote:

The Tao said:
Why are you sterilizing the needle, putting it down then attaching the needle to the syringe?
Attach the needle to the syringe then heat the needle.



I am still learning. I will attach it to the syringe then heat it in future.


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OfflineTakeatrip
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Registered: 09/10/23
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Re: BRF cake contamination cause with Golden Teacher? [Re: BrokenHeart]
    #28637546 - 01/28/24 12:46 AM (7 hours, 13 minutes ago)

Quote:

BrokenHeart said:
The GT syringes I bought for study barely had anything visible. Having many visible spores sounds like a good idea but with the larger spore load the greater the chance for contamination.  In your situation I would use the smallest amount I could on agar (probably 6-10 plates) and then transfer as soon as possible to try to separate good from bad.



I am attempting to do this at the moment. My first syringe had nothing visible but this new one has many spores under a magnifying glass, but as you say I have read in these forums too many spores can cause contamination as some is just inherently in there. I will continue with more petri dishes, but man I have gone through so many its surprising.


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OfflineTakeatrip
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Registered: 09/10/23
Posts: 103
Last seen: 6 hours, 44 minutes
Re: BRF cake contamination cause with Golden Teacher? [Re: LadysKnight]
    #28637549 - 01/28/24 12:58 AM (7 hours, 2 minutes ago)

Quote:

LadysKnight said:
I could be misunderstanding, but it sounds like you flame the needle, let it cool, then attach it to the syringe?

I would attach the needle, shake then flame.

Either way, it looks like the syringe is fucked. I don't think it's you.



I don’t think its me either. I have made a really special effort with my SAB as it has a stainless steel bottom on it now and I have made the hand holes bigger and covered them with gorilla tape on both sides then cut a giant X onto both holes. I have added small magnets with the polarity facing one way right on all 4 flaps near the centre of each hole and then I have cut two small pieces of steel sheet to stick onto the magnets to keep the air inside as still as possible when not in use.
I have gone the extra mile with it because I kept getting contamination with different things. It is such a fascinating hobby, I have learned so much and yet so little at the same time.


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InvisibleLadysKnight
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Re: BRF cake contamination cause with Golden Teacher? [Re: Takeatrip]
    #28637633 - 01/28/24 06:12 AM (1 hour, 48 minutes ago)

Quote:

Takeatrip said:
Quote:

LadysKnight said:
I could be misunderstanding, but it sounds like you flame the needle, let it cool, then attach it to the syringe?

I would attach the needle, shake then flame.

Either way, it looks like the syringe is fucked. I don't think it's you.



I don’t think its me either. I have made a really special effort with my SAB as it has a stainless steel bottom on it now and I have made the hand holes bigger and covered them with gorilla tape on both sides then cut a giant X onto both holes. I have added small magnets with the polarity facing one way right on all 4 flaps near the centre of each hole and then I have cut two small pieces of steel sheet to stick onto the magnets to keep the air inside as still as possible when not in use.
I have gone the extra mile with it because I kept getting contamination with different things. It is such a fascinating hobby, I have learned so much and yet so little at the same time.




Yeah I think it's definitely the syringe. Your SAB sounds good, but if I'm understanding you correctly, it could be improved by removing the gorilla tape. The holes should be 6-7 inches, 15-18cm wide. With the tape, it sounds like you're back to holes that are tight to your arms. You want those holes wide open while you're working. There will be less of a piston effect when you put your hand in or out.


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