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OfflinePhosCap
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Re: AGAR ENVY! (Anything and All things agar!) [Re: Caps McGee]
    #25724246 - 01/05/19 01:27 AM (5 years, 24 days ago)

Isolated?



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InvisibleCaps McGee
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Registered: 10/28/17
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Re: AGAR ENVY! (Anything and All things agar!) [Re: PhosCap]
    #25724661 - 01/05/19 08:36 AM (5 years, 23 days ago)

How'd we get to this point? (MS, clone culture, etc) I don't see any obvious sectoring, but I typically don't see those until I'm close to isolation anyway


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OfflineHumanhelper
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Re: AGAR ENVY! (Anything and All things agar!) [Re: Tormato]
    #25724677 - 01/05/19 08:47 AM (5 years, 23 days ago)

What the absolute feckleberries!?!? Everything is just a little too connected and perfectly lined up today! I am literally, litraly, prepping my workspace and getting ready to weight out some PDA for my first agar pours. I’ve been reading up all week and you sons a bitches are alright!!! Got all the didactic shit outta the way and here goes....then lo and behold, I think to check the ol shroomery and what do I find?! An agar thread at the top of recent posts! Hell ya! I dig!
So, just a thanks from a dude who loves to learn, help other folks when he can, and generally wants everybody to just be okay!
HH


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Offlinemushroomnate
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Re: AGAR ENVY! (Anything and All things agar!) [Re: Humanhelper]
    #25724950 - 01/05/19 10:56 AM (5 years, 23 days ago)

Got germ(I think :shrug2:)on some Pan cyan goliath & jamiacan.


Cluster bombed this Galindoi wedge on to this plate.Looks decent,I may still take it to grain.:shrug:


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BRF to bulk/Cakes as spawn


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OfflineShaperDreaming
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Re: AGAR ENVY! (Anything and All things agar!) [Re: mushroomnate]
    #25725572 - 01/05/19 04:46 PM (5 years, 23 days ago)


So I just poured these at the lowest temp yet (118f) and got more condensation even... I'm suspecting it's the radiators in my house now.

Anyways! I've got fun stuff going on. From left to right:
  • Pan Tropicalis - India (used inoculation loop from syringe)
  • Rusty White transfer 2 (I also used the same plate to knock up a jar just because I'm impatient)
  • Penis Envy, transfer 4, strong rhyzomorphic growth, final transfer before the drop.
  • APE transfer 2 (the plate it came from was the swab plate, so it was challenging to find a good spot to transfer from, still needs a lot of cleaning up).
  • Golden Teacher clone transfer 2 (small isolate area was slightly stronger looking, mostly transfered to clean up the plate)
  • Pear Oyster, print scrape (should be interesting since the spores were white and basically invisible)
  • Golden Oyster, Attempt 2, Transfer 2 (cleaning this one up, but it was looking pretty clean before transfer).


So looks like I need some straw soon ish!

Also, anyone know a good pan tropicalis tek? I have no clue what I'm doing with this one.


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OfflinePhosCap
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Re: AGAR ENVY! (Anything and All things agar!) [Re: Caps McGee]
    #25725696 - 01/05/19 05:37 PM (5 years, 23 days ago)

Quote:

Caps McGee said:
How'd we get to this point? (MS, clone culture, etc) I don't see any obvious sectoring, but I typically don't see those until I'm close to isolation anyway




Started out from spore. I believe it is 3rd transfer. Tried to cut as little as possible. I thought a concentric ring is a sign its isolated.?


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InvisibleCaps McGee
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Re: AGAR ENVY! (Anything and All things agar!) [Re: PhosCap]
    #25725770 - 01/05/19 06:09 PM (5 years, 23 days ago)

No... it's a sign the culture is sensitive to light cycle or temperature fluctuations... you're likely many, MANY transfers away from isolation... you'll eventually notice sectoring, like different cultures in pie slices from the center out... then you're likely still 5-8 transfers from isolating a single strain... I suggest running MS tubs as genetically variant as possible, picking a cluster fruit to clone and further isolate from(as even the culture from a single fruit will contain multiple strains)... you could make a hundred transfers to isolate, and end up with a culture that performs poorly, is low in actives, or even absolutely will not fruit... why are you isolating from spore?


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OfflinePhosCap
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Re: AGAR ENVY! (Anything and All things agar!) [Re: Caps McGee]
    #25725881 - 01/05/19 07:09 PM (5 years, 23 days ago)

Quote:

Caps McGee said:
No... it's a sign the culture is sensitive to light cycle or temperature fluctuations... you're likely many, MANY transfers away from isolation... you'll eventually notice sectoring, like different cultures in pie slices from the center out... then you're likely still 5-8 transfers from isolating a single strain... I suggest running MS tubs as genetically variant as possible, picking a cluster fruit to clone and further isolate from(as even the culture from a single fruit will contain multiple strains)... you could make a hundred transfers to isolate, and end up with a culture that performs poorly, is low in actives, or even absolutely will not fruit... why are you isolating from spore?




ah ok.. i need to remember this. I think I read a couple years ago about this. Ive been stopping and culturing then I forget these things. Ok so next round will be taking some tissue from a nice cluster.

Why can't you isolate from spore? Will it take forever to find an isolate?

I have some somewhat old mush that have grown out from my agar. I should get those tissue to agar perhaps.


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InvisibleCaps McGee
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Registered: 10/28/17
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Re: AGAR ENVY! (Anything and All things agar!) [Re: Caps McGee]
    #25725907 - 01/05/19 07:24 PM (5 years, 23 days ago)

You absolutely can isolate from spore, but it will take forever without lab equipment, and like I stated in the comment you quoted and responded to:

Quote:

Caps McGee said:
No... it's a sign the culture is sensitive to light cycle or temperature fluctuations... you're likely many, MANY transfers away from isolation... you'll eventually notice sectoring, like different cultures in pie slices from the center out... then you're likely still 5-8 transfers from isolating a single strain... I suggest running MS tubs as genetically variant as possible, picking a cluster fruit to clone and further isolate from(as even the culture from a single fruit will contain multiple strains)... you could make a hundred transfers to isolate, and end up with a culture that performs poorly, is low in actives, or even absolutely will not fruit... why are you isolating from spore?




There's no way of knowing what you're isolating until it fruits... IF it fruits at all...

Waste a lot of time and resources on essentially crap


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OfflineHumanhelper
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Re: AGAR ENVY! (Anything and All things agar!) [Re: Tormato]
    #25726732 - 01/06/19 08:58 AM (5 years, 22 days ago)

Noob with a question.  Is this amount of condensation acceptable/normal? PDA, 12g in 250mL, PC’d with filter patch vent in mason jar, vented 15 minutes, 15psi 15minnutes, poured approximately 1100 hours in SAB using normal lab pour technique to sterile 100x15 dishes with Parafilm applied. Specimen just so happened to break veil at 2000 same day so I was forced to clone and print.  Only three samples taken to dishes seen in pics. This morning room temp is 79F and I notice quite a bit of rivulets of H2O in media and condensation on lids. Looks good? Or too wet? Love the feedback!
HH



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OfflineDuhrwood Stevens
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Re: AGAR ENVY! (Anything and All things agar!) [Re: Humanhelper]
    #25726738 - 01/06/19 09:03 AM (5 years, 22 days ago)

Did you pour in stacks starting at the bottom and leave stacked to cool? Usually the stacked ones don't steam up except the top.


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OfflineBeefSupremeJr
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Re: AGAR ENVY! (Anything and All things agar!) [Re: Duhrwood Stevens]
    #25726773 - 01/06/19 09:31 AM (5 years, 22 days ago)

Myco-humans.  Take this for what it’s worth coming from a Hort-human:

In my 17 years of nursery experience I have dealt with condensation in a lot of different ways,  the thing is, water vapor will always condense on whatever is coldest.  That’s why the bottom of the stack tends not to condense—they’re insulated. 

I used to wear jewelers glasses to do much of my seeding work,  I kept them on the heating pads so they wouldn’t fog up from being cold. 

Whenever someone with glasses was about to walk into the greenhouse, I’d always tell them to take their glasses off and put them in their pocket to warm up a bit. Otherwise they immediately fog up the moment they walked in and would be disoriented.

The plastic domes over my seedling trays would condense all the time.  There wasn’t much that could be done for it.  When big droplets started to form, I’d shake them out before replacing the lid to avoid big water bombs from damaging the top dress.

The galvanized rails that suspended our lights tend to condense water and drip water bombs onto cutting trays.  The solution was to insulate them. 

How does that apply to plates? Beats me.  The lids are colder than the agar so of course they will condense.  That’s physics.  Cooler agar would help a little I suppose.  Hotter lids would be the real ticket. How to do that?  I thought about a heat pad in the sab, but boy that would create a convection cycle and totally ruin the still air environment, but perhaps no more than the hot bottle of agar itself.

That’s all I can tell you.  The lids must be heated to avoid condensation and allowed to cool in tandem with the agar itself.  That’s just plain physics. 

How to go about doing that is another story.  A hot sab would do it,  but I think you’re asking for more problems than it’s worth.


Edited by BeefSupremeJr (01/06/19 09:33 AM)


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OfflineHumanhelper
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Re: AGAR ENVY! (Anything and All things agar!) [Re: Duhrwood Stevens]
    #25726910 - 01/06/19 10:41 AM (5 years, 22 days ago)

Quote:

Duhrwood Stevens said:
Did you pour in stacks starting at the bottom and leave stacked to cool? Usually the stacked ones don't steam up except the top.



Yes, I did. Just like I would in the lab in front of a flow hood. My thoughts are maybe too much water in mix? OR, I didn’t let em sit long enough after a pour. In the lab the plates are poured and allowed to sit for 3-5 days to verify no contams. I didn’t have time for that since I had big daddies poppin, lol.


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OfflineHumanhelper
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Re: AGAR ENVY! (Anything and All things agar!) [Re: BeefSupremeJr]
    #25726920 - 01/06/19 10:45 AM (5 years, 22 days ago)

Quote:

BeefSupremeJr said:
Myco-humans.  Take this for what it’s worth coming from a Hort-human:

In my 17 years of nursery experience I have dealt with condensation in a lot of different ways,  the thing is, water vapor will always condense on whatever is coldest.  That’s why the bottom of the stack tends not to condense—they’re insulated. 

I used to wear jewelers glasses to do much of my seeding work,  I kept them on the heating pads so they wouldn’t fog up from being cold. 

Whenever someone with glasses was about to walk into the greenhouse, I’d always tell them to take their glasses off and put them in their pocket to warm up a bit. Otherwise they immediately fog up the moment they walked in and would be disoriented.

The plastic domes over my seedling trays would condense all the time.  There wasn’t much that could be done for it.  When big droplets started to form, I’d shake them out before replacing the lid to avoid big water bombs from damaging the top dress.

The galvanized rails that suspended our lights tend to condense water and drip water bombs onto cutting trays.  The solution was to insulate them. 

How does that apply to plates? Beats me.  The lids are colder than the agar so of course they will condense.  That’s physics.  Cooler agar would help a little I suppose.  Hotter lids would be the real ticket. How to do that?  I thought about a heat pad in the sab, but boy that would create a convection cycle and totally ruin the still air environment, but perhaps no more than the hot bottle of agar itself.

That’s all I can tell you.  The lids must be heated to avoid condensation and allowed to cool in tandem with the agar itself.  That’s just plain physics. 

How to go about doing that is another story.  A hot sab would do it,  but I think you’re asking for more problems than it’s worth.



Thanks for sharing your thoughts. I have a pretty solid grasp of the physics, but probably more the chemistry of it all from my other life, but my main concern is the biology of it all:). Will the excess water harm/hamper the growth of the specimen?  Also, not being a know-it-all, just agreeing with you on the physics. This is literally the greats mind expanding hobby on every layer!
HH


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OfflineDuhrwood Stevens
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Re: AGAR ENVY! (Anything and All things agar!) [Re: Humanhelper]
    #25726929 - 01/06/19 10:51 AM (5 years, 22 days ago)

Not too much water, too hot when poured, likely. Doesn't really matter. Hard to see is the worst part. Pour hot stuff in a cool room it sweats.
Check this 'fix' from Bod.
https://www.shroomery.org/forums/showflat.php/Number/21922023
''how do you fix them?

use a cup with warm water and a flat bottom, put it on top of the dish for a few seconds-minutes (wont hurt if the water is warm-hot)
and boom your dishes are crystal clear again.''


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OfflineBeefSupremeJr
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Re: AGAR ENVY! (Anything and All things agar!) [Re: Humanhelper]
    #25726933 - 01/06/19 10:55 AM (5 years, 22 days ago)

I hear you my dude.  I’m not trying to act like a know it all either, just saying: this is what’s happening and why it’s happening. 

How to stop it from happening is another story.  Hoping caps or bodhi figure something out eventually. 

As far as condensation causing issues, it doesn’t.  At all.  Its just that clear plates are so pretty.



I thought about trying to take a real foggy plate, microwaving it for just long enough to liquify the agar and see if it helps.  Big maybe.  Probably ruin the plate, but I’ll sure let you know how it goes.


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OfflineBeefSupremeJr
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Re: AGAR ENVY! (Anything and All things agar!) [Re: Duhrwood Stevens]
    #25726935 - 01/06/19 10:56 AM (5 years, 22 days ago)

Quote:

Duhrwood Stevens said:
Not too much water, too hot when poured, likely. Doesn't really matter. Hard to see is the worst part. Pour hot stuff in a cool room it sweats.
Check this 'fix' from Bod.
https://www.shroomery.org/forums/showflat.php/Number/21922023
''how do you fix them?

use a cup with warm water and a flat bottom, put it on top of the dish for a few seconds-minutes (wont hurt if the water is warm-hot)
and boom your dishes are crystal clear again.''





See, it’s probably just that easy.  I suspect that if you poured in a 90degree room, you would t have this issue at all.  But who the fuck wants to do that?


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OfflineHumanhelper
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Re: AGAR ENVY! (Anything and All things agar!) [Re: BeefSupremeJr]
    #25726945 - 01/06/19 11:01 AM (5 years, 22 days ago)

Awesome info! Thanks all! I agree I wanted “pretty” plates, but I’ll take clean over pretty any day! Good to know moisture won’t harm...and I’ll check out bod’s fix..I’ll post as I go, early days still:) holy shit I’m using a lot of smileys and lols. The morning coffee and herb concoction is working out well for me:)
HH


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OfflineBeefSupremeJr
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Re: AGAR ENVY! (Anything and All things agar!) [Re: Humanhelper]
    #25726950 - 01/06/19 11:03 AM (5 years, 22 days ago)

Yeah but those pretty plates are just so fucking pretty.  I love pretty things.


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InvisibleJ. Jack Flash
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Registered: 11/20/13
Posts: 1,500
Re: AGAR ENVY! (Anything and All things agar!) [Re: BeefSupremeJr]
    #25726967 - 01/06/19 11:11 AM (5 years, 22 days ago)

Quote:

BeefSupremeJr said:
See, it’s probably just that easy.  I suspect that if you poured in a 90degree room, you would t have this issue at all.  But who the fuck wants to do that?




hot yoga enthusiasts?

the room would have to be warmer than the agar you're pouring. it's the temperature gradient that causes the condensation.

really tho, cooling them to room temp before covering is how to prevent condensation.

we need to invent the home gamma irradiator. pour plates in open air, cool to room temp, cover, and irradiate to sterility. simple.


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