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InvisibleNigel Stephens
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Registered: 02/21/20
Posts: 103
Re: AGAR ENVY! (Anything and All things agar!) [Re: bigfootscreepyuncl]
    #27551077 - 11/20/21 11:48 AM (2 years, 4 months ago)

Thank-you for the suggestion, I’ll refrain from shitposting to get those 6 remaining. In the meantime, anything I can do with my T1 and germ plates to try and get things moving again?

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InvisibleNigel Stephens
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Registered: 02/21/20
Posts: 103
Re: AGAR ENVY! (Anything and All things agar!) [Re: nektar61]
    #27551086 - 11/20/21 11:52 AM (2 years, 4 months ago)

Quote:

nektar61 said:
Quote:

Nigel Stephens said:
I’m struggling somewhat, and could do with some next step advice please!

A few weeks ago I put some spores from a print to agar (2% LME). The spores are about 18months old.

Only after 3 weeks did I see anything resembling mycelium and by this time the plates were fully grown out with what looks like a white mould...




Clear photos please.

It can take a month for spores to show growth on agar. In my experience it usually takes less, and with fresh spores I've seen it in 3 days, (some have mentioned a day), but we need pix to help.




I will when I’m back with my plates, cheers

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OfflineMarcus666
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Posts: 162
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Re: AGAR ENVY! (Anything and All things agar!) [Re: Mr. Mushie]
    #27551087 - 11/20/21 11:52 AM (2 years, 4 months ago)

This is an example of the plates I took transfers off.



Same recipe, something's gone wonky somewhere along the line.

I'll grab some yeast and give that a go.

Thanks folks!

Edited by Marcus666 (11/20/21 11:54 AM)

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InvisibleNigel Stephens
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Registered: 02/21/20
Posts: 103
Re: AGAR ENVY! (Anything and All things agar!) [Re: Mr. Mushie]
    #27551089 - 11/20/21 11:53 AM (2 years, 4 months ago)

Quote:

Mr. Mushie said:
Quote:

Nigel Stephens said:
I’m struggling somewhat, and could do with some next step advice please!

A few weeks ago I put some spores from a print to agar (2% LME). The spores are about 18months old.

Only after 3 weeks did I see anything resembling mycelium and by this time the plates were fully grown out with what looks like a white mould, forming disorganised and occasionally fluffy white patches. The mould is not dense, and has not produced any other colours by sporulation.

Anyway, from the tendrils of organised looking mycelium (rhizo) which I could spy amongst this stuff I took 2 transfers.

I transferred them to 1.5% LME agar in an effort to stimulate some rhizo leap-off to then transfer away again.

However a week later, nothing has grown from the transfers. Nothing at all. No bacteria, no mould, no contams of any sort and no mycelium.

I have no more spores, my control plates are clean as a whistle.

I thought I could do a hot pour of 2% agar again onto the T1 plates but this is likely flawed. Short of starting again (can’t seem to get spores!) - what would you guys do?

Thanks in advance




PM me, i'd be happy to send you a few different varieties to work with.




You are an absolute hero! Thankyou

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Offlinenektar61S
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Re: AGAR ENVY! (Anything and All things agar!) [Re: Nigel Stephens]
    #27551104 - 11/20/21 12:01 PM (2 years, 4 months ago)

Quote:

Nigel Stephens said:
You are an absolute hero! Thankyou




Most of us don't send addresses through this site, most folks use this
https://privnote.com/

self-destructs after the recipient reads it. Make one and try it. Hit the link twice. first time works, second time doesn't.


--------------------
-NEW? Start here.

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InvisibleNigel Stephens
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Re: AGAR ENVY! (Anything and All things agar!) [Re: Marcus666] * 1
    #27551119 - 11/20/21 12:11 PM (2 years, 4 months ago)

Quote:

Marcus666 said:
Sorry for the external link (if it's not allowed please delete it Mods! and I apologise.)

But is this stuff ok to use ??? "Nut Yeast"...

https://www.hollandandbarrett.com/shop/product/engevita-yeast-flakes-b12-60009142

:aliendance:




Dude I’ve used this in the past and it wasn’t very soluble. Some dissolves sure, but plates had specs in. If that doesn’t bother you, no worries. If it does, consider straining your agar pre PC cycle or maybe put it in a coffee grinder to whizz it up. I did have growth using it so it’s not an inhibitor or anything!

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InvisibleNigel Stephens
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Posts: 103
Re: AGAR ENVY! (Anything and All things agar!) [Re: nektar61]
    #27551121 - 11/20/21 12:13 PM (2 years, 4 months ago)

Quote:

nektar61 said:
Quote:

Nigel Stephens said:
You are an absolute hero! Thankyou




Most of us don't send addresses through this site, most folks use this
https://privnote.com/

self-destructs after the recipient reads it. Make one and try it. Hit the link twice. first time works, second time doesn't.




Very helpful thanks

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OfflineDefacto2
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Last seen: 1 year, 10 months
Re: AGAR ENVY! (Anything and All things agar!) [Re: nektar61]
    #27551214 - 11/20/21 01:48 PM (2 years, 4 months ago)

Thanks for the link & tip!

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InvisibleGuerrilla
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Re: AGAR ENVY! (Anything and All things agar!) [Re: Nigel Stephens]
    #27551238 - 11/20/21 02:06 PM (2 years, 4 months ago)

Quote:

Nigel Stephens said:
Quote:

Marcus666 said:
Sorry for the external link (if it's not allowed please delete it Mods! and I apologise.)

But is this stuff ok to use ??? "Nut Yeast"...

https://www.hollandandbarrett.com/shop/product/engevita-yeast-flakes-b12-60009142

:aliendance:




Dude I’ve used this in the past and it wasn’t very soluble. Some dissolves sure, but plates had specs in. If that doesn’t bother you, no worries. If it does, consider straining your agar pre PC cycle or maybe put it in a coffee grinder to whizz it up. I did have growth using it so it’s not an inhibitor or anything!




Yeah yeast doesn't dissolve, no brand does. You filter it through coffee filters if you're bothered about the sediment.

Yeast in pot of water, boil, strain, add agar, back into the pot, then to media bottles and PC. This works but you don't want to strain your whole agar solution pre PC because you'll be filtering out the agar which will collect in the filter.


--------------------
Being pissed on does not make you a real man.

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Offlinenektar61S
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Re: AGAR ENVY! (Anything and All things agar!) [Re: Guerrilla]
    #27551324 - 11/20/21 03:08 PM (2 years, 4 months ago)

LC a la Mushboy.
4 bottles. About 150 ML each.
--T1 (stipe clone) PE6
--T1 (swab clone) Penis Envy
--T1 (swab clone) Alacabenzi
--T4 (spores) RustyWhyte


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Offlinem0n0id
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Re: AGAR ENVY! (Anything and All things agar!) [Re: nektar61]
    #27551384 - 11/20/21 04:01 PM (2 years, 4 months ago)

I too have been having some trouble with getting desirable growth, though only having trouble with germination (cubes, pan cyan)...

One other factor in getting lackluster growth could be sterilization time/pressure. This might be applicable to Nigel as well...

What I've done...

In both of the following cases I was running for 45m, which I'm gathering is way too long.
To start I was using a leaky old ass PC and running it at 15psi. Had no trouble germinating. Then I got an AA sterilizer and started running 18psi (did I mention I suck at following directions?), and in addition spending a lot more time doing AA's recommended vent after reaching pressure. (I was using the AA sterilizer insert.)

Been using a premixed MEYA which I think is about so... (Kinda guessing based on the dude's product label and book, you can probably guess who I'm talking about.)
For 1000ml water:
20g agar
25g malt extract
5g yeast

Semi-scientific speculation

Did a little reading about caramelization on wikipedia today... Turns out parallelization is polymerization of sugars, which in layman's terms means bonding smaller molecules together into big ones. Guessing that the bigger caramel molecules are harder for mycelium to eat compared to simpler sugar molecules.

What I plan to do...

Sterilization time: 20m @ 15psi
AA Sterilizer run in PC mode
  No inset this time, vent hose tied in a knot to keep it well above water
  Media bottles partially immersed for good heat conduction

Maybe add a couple grams of peptone?

Any other thoughts or recommendations?

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Offlinenektar61S
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Re: AGAR ENVY! (Anything and All things agar!) [Re: m0n0id]
    #27551425 - 11/20/21 04:31 PM (2 years, 4 months ago)

Quote:

m0n0id said:
I too have been having some trouble with getting desirable growth, ....
Any other thoughts or recommendations?




I would skip the pan cyan until you have decent success with a cube variety.

Most people here are not using peptone. A few are, but not most. It's certainly not the missing ingredient in not getting contam. 

What's a vent hose?

Are you screwing the media bottles shut? That keeps the steam out. I put the lid barely on, very loose. Then wrap aluminium foil loosely on them.

This agar mix is working very well for me:
https://www.shroomery.org/forums/showflat.php/Number/27551422/vc/1#27551422


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Edited by nektar61 (11/20/21 04:46 PM)

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Offlinem0n0id
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Re: AGAR ENVY! (Anything and All things agar!) [Re: nektar61]
    #27551457 - 11/20/21 04:59 PM (2 years, 4 months ago)

I've had success with cubes, just relatively shy about posting pics and progress. Definitely a little on the paranoid side. Definitely got plenty to learn still!

Yeah, peptone is definitely not the missing key for me; had good success with MEYA on cubes and isolated a strain that I'm pretty happy with.

On the AA sterilizer models (not PC) there is an inner insert which is kinda like a cooking pot. From the vent valve on the lid a tube dangles down to the bottom of the insert. The point of this is to allow the steam from the water in the bottom to come up around the sides of the insert down into the top of the insert and push the cold air pooling at the bottom out through the vent hose. The AA sterilizer vent tube design is kinda a siphon for the air (nitrogen, oxygen, CO2, etc.) which presumably settles in the bottom of the insert.

Sorry that's a little long winded... haha!

Thanks for the link! I've still got a bunch of premixed MEYA to work through, but I'll keep it it mind for when that's gone.

Quick edit: Yep, I'm already doing what you're suggesting with keeping the media bottles cracked and foiling them.

Edited by m0n0id (11/20/21 05:00 PM)

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InvisibleWyoMX
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Registered: 07/06/15
Posts: 2,193
Loc: PNW
Re: AGAR ENVY! (Anything and All things agar!) [Re: m0n0id] * 1
    #27551467 - 11/20/21 05:07 PM (2 years, 4 months ago)

One thing I do like to point out to newer users shy with pics is that this site strips all exif data from pics. So it's a little better but definitely don't show any identifying features. Definitely understand your concern though and won't push one way or another. As for the agar issues I would recommend to not use the premix stuff. I only tried 2 different brands of it but didn't have nearly as much success as when I mix my own malt extract and agar powder.  When I say success I basically mean that all my cultures just liked my recipes better then the premix.

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Offlinenektar61S
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Re: AGAR ENVY! (Anything and All things agar!) [Re: m0n0id]
    #27551474 - 11/20/21 05:12 PM (2 years, 4 months ago)

Quote:

m0n0id said:
I've had success with cubes, just relatively shy about posting pics and progress. Definitely a little on the paranoid side. Definitely got plenty to learn still!

Yeah, peptone is definitely not the missing key for me; had good success with MEYA on cubes and isolated a strain that I'm pretty happy with.

On the AA sterilizer models (not PC) there is an inner insert which is kinda like a cooking pot. From the vent valve on the lid a tube dangles down to the bottom of the insert. The point of this is to allow the steam from the water in the bottom to come up around the sides of the insert down into the top of the insert and push the cold air pooling at the bottom out through the vent hose. The AA sterilizer vent tube design is kinda a siphon for the air (nitrogen, oxygen, CO2, etc.) which presumably settles in the bottom of the insert.

Sorry that's a little long winded... haha!

Thanks for the link! I've still got a bunch of premixed MEYA to work through, but I'll keep it it mind for when that's gone.

Quick edit: Yep, I'm already doing what you're suggesting with keeping the media bottles cracked and foiling them.




Are you letting the plates sit taped up 7-10 days to check for contams? that was a game changer for me.


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Offlinem0n0id
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Re: AGAR ENVY! (Anything and All things agar!) [Re: WyoMX] * 1
    #27551492 - 11/20/21 05:32 PM (2 years, 4 months ago)

@WyoMX

Thanks for pointing the EXIF stripping feature out, that's a really nice feature to have. I'll probably still strip  EXIF on my end before posting, but I'll get over the paranoia at some point.

The fact that Shroomery is still here and people repeatedly posting large volume cult results makes it pretty obvious that it is relatively safe.


@nektar61

Yeah, I've done that as well, using parafilm, haven't really had too much issues with contams. A bit of bacteria here and there and no mold whatsoever. I'm working with a flowhood BTW.

Wondering if my issues might be related to spore application technique? Here's two failed instances:

Cubes: I used an inoculation loop that I'd flamed and cooled in the recipient dish (which had literally set for months without contam). Rubbed it on a fairly thin sporeprint from a previous grow. Had no success there; thinking the issue may have been that the spores were stuck too well to the foil, but that doesn't really pass the smell test given how numerous spores are on a print.

Pans: Got a syringe from a vendor. Was an idiot and squirted way too much (several drops) onto the dish and streaked it. This did contam, and I've no one to blame but myself. Taking a second try with pans, where I dripped a drop from the syringe onto the loop, flicked the drop off the loop and then streaked. Hoping this works out.

Oysters: I remember my first try streaking spores didn't work but then after dipping the loop in aseptic water and then sampling the print it took off just fine. I will try this technique next time I try to cult cubes from my prints.

The starter print I got from a FOAF was from a very mature fruit so it was layered thick and not stuck to the foil. I've probably been printing relatively immature fruits which is why I've been getting thin spore depositions that really stick hard to the foil.


@Everyone
Gotta say a big thank you to all of the folks on here posting great info! It is very very helpful!

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InvisibleGuerrilla
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Re: AGAR ENVY! (Anything and All things agar!) [Re: nektar61]
    #27551496 - 11/20/21 05:35 PM (2 years, 4 months ago)

Quote:

nektar61 said:
Quote:

m0n0id said:
I too have been having some trouble with getting desirable growth, ....
Any other thoughts or recommendations?




I would skip the pan cyan until you have decent success with a cube variety.[/url]




I know a few people that have only grown pans and never cubes. Baba Yaga's pans in monotubs might be helpful.


--------------------
Being pissed on does not make you a real man.

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Offlinem0n0id
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Re: AGAR ENVY! (Anything and All things agar!) [Re: Guerrilla]
    #27551517 - 11/20/21 05:55 PM (2 years, 4 months ago)

Quote:

Guerrilla said:I know a few people that have only grown pans and never cubes. Baba Yaga's pans in monotubs might be helpful.




That's definitely an interesting one, looks like Baba Yaga has also had some decent success with coir only which is pretty wild. I've been reading heavily from the movers on the Cope/Pan Thread, lots of great info.

Pans are appealing for being able to get a high dose with less fungus flesh. Also looking for a bit of a challenge. I found that cubes nearly grew themselves on coir. I got freaked out cause of the quantity I was producing and just neglected the coir after one flush and they went on to pop out some massive shrooms despite my best efforts to dry them out to kill em.



As for my current germination woes I'm sure I'll get past em by experimenting. It's worked in the past :wink:

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OfflineDERRAYLD
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Re: AGAR ENVY! (Anything and All things agar!) [Re: m0n0id] * 2
    #27551880 - 11/21/21 12:53 AM (2 years, 4 months ago)

Liking the look of this T1, very clean and uniform currently.


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InvisibleD3_Myc
Weeb Trash
I'm a teapot


Registered: 05/06/18
Posts: 4,399
Loc: Year Zero
Trusted Cultivator
Re: AGAR ENVY! (Anything and All things agar!) [Re: DERRAYLD]
    #27551888 - 11/21/21 01:02 AM (2 years, 4 months ago)

Quote:

DERRAYLD said:
Liking the look of this T1, very clean and uniform currently.







Look forward to seeing what it does for you!

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