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Offlinerumfor69
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Re: AGAR ENVY! (Anything and All things agar!) [Re: Mr Piggy] * 4
    #28774045 - 05/14/24 11:31 AM (1 month, 11 days ago)

Go with the CS method! Just need sterile coverslips

https://pubmed.ncbi.nlm.nih.gov/31693673/


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InvisibleYuggoth
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Re: AGAR ENVY! (Anything and All things agar!) [Re: rumfor69] * 1
    #28774054 - 05/14/24 11:50 AM (1 month, 11 days ago)

Quote:

rumfor69 said:
Go with the CS method! Just need sterile coverslips

https://pubmed.ncbi.nlm.nih.gov/31693673/




Now this is cool as shit.


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InvisibleLadysKnight
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Re: AGAR ENVY! (Anything and All things agar!) [Re: rumfor69]
    #28774061 - 05/14/24 11:58 AM (1 month, 11 days ago)

Quote:

rumfor69 said:
Go with the CS method! Just need sterile coverslips

https://pubmed.ncbi.nlm.nih.gov/31693673/




Nice. Looks like sandwich tek but with a coverslip. The advantage of the sandwich tek would be the ability of myc to also grow through the cover piece.

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OfflineAspectOfTheCreator
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Re: AGAR ENVY! (Anything and All things agar!) [Re: mycosispeon]
    #28774073 - 05/14/24 12:16 PM (1 month, 11 days ago)

I'm so used to jars only taking 2.5 weeks to colonize, I'll exercise more patience though and spawn them once the time comes.


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Offlinerumfor69
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Re: AGAR ENVY! (Anything and All things agar!) [Re: LadysKnight]
    #28774091 - 05/14/24 12:43 PM (1 month, 11 days ago)

Quote:

LadysKnight said:
Quote:

rumfor69 said:
Go with the CS method! Just need sterile coverslips

https://pubmed.ncbi.nlm.nih.gov/31693673/




Nice. Looks like sandwich tek but with a coverslip. The advantage of the sandwich tek would be the ability of myc to also grow through the cover piece.




You maybe could use a larger piece of agar as a coverslip. What I find interesting is "the cabin" technique.


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Offlinerumfor69
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Re: AGAR ENVY! (Anything and All things agar!) [Re: rumfor69] * 1
    #28774097 - 05/14/24 12:53 PM (1 month, 11 days ago)

I wonder though if the coverslip being non nutritious is the key to keeping bacteria trapped in "the cabin". Probably could make your own coverslips outta silicone and sterilize them in an empty dish then just transfer it over your cabin you placed the bacterial enmeshed transfer in. The mycelium reaches out has to cross the space of the cabin then squeeze out in-between the coverslip and agar and leaves bacteria trapped in the cabin.


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InvisibleMr Piggy
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Re: AGAR ENVY! (Anything and All things agar!) [Re: rumfor69]
    #28774111 - 05/14/24 01:03 PM (1 month, 11 days ago)

Quote:

rumfor69 said:
Go with the CS method! Just need sterile coverslips

https://pubmed.ncbi.nlm.nih.gov/31693673/




That's cool and all, but I'm talking about trich and not bacteria.  I'm a lot lazier than a lot of y'all, I don't fight for what doesn't fight on it's own :shrug:


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Offlinerumfor69
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Re: AGAR ENVY! (Anything and All things agar!) [Re: Mr Piggy] * 2
    #28774126 - 05/14/24 01:21 PM (1 month, 11 days ago)

I've never seen enmeshed mold on agar and I doubt trich can even do that.

But enmeshed bacteria is a very real thing. I've delt with a few times trying to save favorite cultures. That CS method was developed purposely to get away from unseen bacteria that can't just be transferred away from.


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Re: AGAR ENVY! (Anything and All things agar!) [Re: rumfor69] * 1
    #28774273 - 05/14/24 03:52 PM (1 month, 11 days ago)

T1 HBP clone, definitely going to back up this culture in a cyro vial before putting to grain.

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Offlineunablefrog
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Re: AGAR ENVY! (Anything and All things agar!) [Re: Psilosion] * 8
    #28774354 - 05/14/24 05:05 PM (1 month, 11 days ago)


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Offlinetittlez
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Re: AGAR ENVY! (Anything and All things agar!) [Re: rumfor69]
    #28774815 - 05/15/24 05:04 AM (1 month, 11 days ago)

Quote:

rumfor69 said:
Quote:

LadysKnight said:
Quote:

rumfor69 said:
Go with the CS method! Just need sterile coverslips

https://pubmed.ncbi.nlm.nih.gov/31693673/




Nice. Looks like sandwich tek but with a coverslip. The advantage of the sandwich tek would be the ability of myc to also grow through the cover piece.




You maybe could use a larger piece of agar as a coverslip. What I find interesting is "the cabin" technique.




From what i understand, "the cabin" is simply a small hole in the agar surface in which to place the contaminated mycelium. I guess this should  help in placing the cover slip on top in a way that it seals well, making sure the mycelium must grow through the agar medium.

Indeed this sounds exactly like the sandwich tek to me - only with the coverslip instead. I wonder what is easier when it comes to handling and how exactly the small round or square cabin is made? These to me were the most tricky parts when i attempted the Sandwich TEK a while back. I almost imagine it to be easier to use the sandwich tek seeing as sterilizing and then handling a sterile cover slip sounds like a job for a surgeon with a lot of patience to me :grin:


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Offlinerumfor69
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Re: AGAR ENVY! (Anything and All things agar!) [Re: tittlez] * 1
    #28774842 - 05/15/24 06:04 AM (1 month, 11 days ago)

You would just cut out like a 5mm(1/4") square and remove it from the receiver plate. Then you would place a tiny piece of agar with your culture on it in the center of the square. You could sterilize a glass slipcover on a small tinfoil base, inside an empty petri dish. Grab it with sterile forceps and place it to cover the whole empty area. Slip covers come in lots of different sizes and you could change all these dimensions to fit whatever size piece if agar and tissue your able to transfer but smaller will probably be better for this method.

It's not really like the sandwich method at all because that uses nutrients against nutrients and is an attempt to get mycelium to grow through it and hope that embedded contamination didn't get carried with it.


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Offlinetittlez
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Re: AGAR ENVY! (Anything and All things agar!) [Re: rumfor69]
    #28774859 - 05/15/24 06:20 AM (1 month, 11 days ago)

Sounds like a good method! I guess i´ll just have to try it out to see what works best for me. Thanks for the feedback :chugbeer:


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InvisibleScemilope
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Re: AGAR ENVY! (Anything and All things agar!) [Re: tittlez] * 4
    #28774931 - 05/15/24 08:45 AM (1 month, 11 days ago)


x7x+ T2


10 months old cryo vial sample. Cube(snape), 3rd day after extraction.

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OfflinetHeMaGe
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Re: AGAR ENVY! (Anything and All things agar!) [Re: Scemilope] * 3
    #28775041 - 05/15/24 10:53 AM (1 month, 10 days ago)

Some F+ from a kind and generous Shroomery member starting to germ and a couple nat t1 plates ready to send.




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OfflineChaos4142
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Re: AGAR ENVY! (Anything and All things agar!) [Re: Scemilope] * 1
    #28775048 - 05/15/24 11:00 AM (1 month, 10 days ago)

Hi folks,

have returned to shroomery after a hiatus of almost 4 years, please let me know if this is the right thread to ask this question.
I saved some spores from my last grow circa 2020, and I thought I was set for life as far as the shelf life of the spores is concerned. In April this year I took them to Agar with no results, made a spore syringe no result, applied pressure to syringe and let them sit for 4 or 5 days, and then knocked up some plates. Meanwhile I sent an SOS on the forums, and some generous members have sent some fresh spore prints which are on there way.(they have to cross borders and snail mail in my part of the world can be dodgy and delayed as fuck)

Meanwhile, one of the plates knocked up with pressure treated spore syringe developed some growth. I did a couple of transfers and the results pics are below. one of them is T1 and another T2. 2 pics each. Apologies for the shitty pics as I am using these PP5 food containers and it is a pain to get correct focus and avoid reflection.



Few things to take into consideration
- I have shifted countries and where I am at it is particularly painful to get some items which are *very* easy to get elsewhere (read downunder or US). I had to ask a family member to carry Agar powder from UK (he was like wtf why do you need a jello making ingredient) So most of my SAB, pressure cooker, micropore tape etc etc is quite ghetto to begin with. But I feel it does its job, my control plates did not show an contam, and its been 6 weeks now.

- My method to make spore syringe was quite rudimentary and although it was done in SAB i have zero doubt that i must have introduced bacteria as well as some skin yeast

- Germination plate did similar tomentose growth (or bacterial super imposed myceilum ?) anyways I transfered the most filamenty look leading edges to T1 (29/4/24) and then same for T2 (6/5/24).

- A few of my transfers became bacterial and were chucked in the bin, I dont have pics but this was like slimey stuff spread around the plate (as those plates had exess moisture roaming around the plate. few bumps working in the smallish SAB and overall poor technique.

- Just as this wasnt enough, the temperatures as of 4/5/24 have climbed to well above 30 Celsius, typical day for last 2 weeks has been Max: 38 Celsius Min: 20 Celsius (yeah Air conditioning is out of question right now). Hence you see these nice rings.
- That dodgy orange colour is from the food coloring, yes I am sure. I have knocked up some grain water plates as well I will come back with proof later.

- T1 plate will show some gaps, thats where the T2 inoculations were taken, this plate didnt develop bacteria or mold (note to self, do transfers while baked)

Okay now since I dont have any other source of spores and I really want to salvage something out of this, Are these plates totally doomed or is there a particular way I can get something to fruit to clone and so on.

Not to be dettered by these setbacks I have done some transfers yesterday to BRF pucks (fingers crossed) and actually did the PP5 PF tek directly from the homemade spore syringe.

phew that was long, I needed to vent I guess
thanks for reading, looking forward to customary support of this amazing community.

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InvisibleNichrome
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Re: AGAR ENVY! (Anything and All things agar!) [Re: Chaos4142]
    #28775174 - 05/15/24 12:44 PM (1 month, 10 days ago)

That looks like woodlover myc. Looks a lot like some ovoid cultures I've seen. Also looks like some tamp cultures. Very odd growth and pigmented mycelium if it is cube...


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OfflineChaos4142
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Re: AGAR ENVY! (Anything and All things agar!) [Re: Nichrome]
    #28775197 - 05/15/24 01:11 PM (1 month, 10 days ago)

Quote:

Nichrome said:
That looks like woodlover myc. Looks a lot like some ovoid cultures I've seen. Also looks like some tamp cultures. Very odd growth and pigmented mycelium if it is cube...




I wish, but it is from B+ spores, meshed together a few prints i had from 3-4 years ago, stored in a drawer exposed to 35 deg C to 2 Deg C.

I may as well knock a millet jar tonight, see if it spread better on grain.

One very important detail i forgot, the agar is potato water, white sugar i think 10g for 500 ml. LME not available here.... lets see if it does anything different on grain water....

I am thinking i put the new plates in an esky with some cold packs to better maintain temperture below 25 C

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InvisibleNichrome
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Re: AGAR ENVY! (Anything and All things agar!) [Re: Chaos4142]
    #28775200 - 05/15/24 01:13 PM (1 month, 10 days ago)

Ahhh yeah the nutrients and additives could equate to off looking patterns.


--------------------
question everything, it's very responsive



“Drink your tea slowly and reverently, as if it is the axis on which the world earth revolves - slowly, evenly, without rushing toward the future.”
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OfflineChaos4142
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Re: AGAR ENVY! (Anything and All things agar!) [Re: Nichrome]
    #28775210 - 05/15/24 01:24 PM (1 month, 10 days ago)

@nichrome Excuse my noobness but they are not 100% bacterial in your view?

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