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OfflineKenetic
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Re: Let's All Grow Mushrooms 2017 [Re: Tookitooki] * 1
    #24027840 - 01/21/17 10:41 AM (7 years, 9 days ago)

I just whisper to the plates very softly....


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Re: Let's All Grow Mushrooms 2017 [Re: Tookitooki]
    #24027845 - 01/21/17 10:43 AM (7 years, 9 days ago)

good stuff. I did regular no-pour plates, swiped the cyan spores from the print with the inoc loop and then poured hot agar over the spores... but I got nothing after two weeks.

I'm going to try spores to lme broth next time and if I see germination this way I plan on dipping the inoc loop in the lc and put it on agar to clean it.


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OfflineKenetic
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Re: Let's All Grow Mushrooms 2017 [Re: Josex]
    #24027859 - 01/21/17 10:49 AM (7 years, 9 days ago)

Seems like pouring hot agar over spores is counterproductive


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Re: Let's All Grow Mushrooms 2017 [Re: Kenetic]
    #24027875 - 01/21/17 10:54 AM (7 years, 9 days ago)

Explain thyself :lol: I did that after stumbling upon one of RR's posts a bit ago when I was researching how to germinate stubborn spores.

Apparently spores can withstand agar at 122f for a short time. If they germinate they'll do it under the layer of poured agar and will eventually resurface leaving any possible bacteria behind.


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Re: Let's All Grow Mushrooms 2017 [Re: Kenetic]
    #24027879 - 01/21/17 10:55 AM (7 years, 9 days ago)

Warm is fine

Agar solidifies at 107f or there abouts. I pour my agar at 116-117 to minimize condensation

I would never pour plates with agar above 130F or so condensation city


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OfflineKenetic
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Re: Let's All Grow Mushrooms 2017 [Re: bodhisatta]
    #24027995 - 01/21/17 11:27 AM (7 years, 9 days ago)

I thought spores start dying around 105 f or so.  Thats about the extent of my reasoning


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Re: Let's All Grow Mushrooms 2017 [Re: Kenetic]
    #24028012 - 01/21/17 11:33 AM (7 years, 9 days ago)

The cloest i've done to this is drop spores a few minuets are pouring and just starting to set.
I didnt notoce a difference but ive only done it like once


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InvisibleTookitooki
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Re: Let's All Grow Mushrooms 2017 [Re: Intelligentxfruit]
    #24028039 - 01/21/17 11:46 AM (7 years, 9 days ago)

From what Im learning, spores are like seeds.  They have an external membrane or something that acts as one.  The same reason we don't use 90% ISO, is the same reason we might try a LC if they don't germinate on normal agar.  They require a moisture content to permeate it's membrane, to get to intake any kind of substance.

The older they are it seems like that membrane really dries up, making it harder to get thru.  I'm sure heat, to an extent, helps the moisture get thru that membrane. 

Please correct me if I'm wrong.  And I'm sure this doesn't apply to everything.  But when talking cubes, this is my collective from all the info I have read


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InvisibleTookitooki
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Re: Let's All Grow Mushrooms 2017 [Re: Josex]
    #24028079 - 01/21/17 12:06 PM (7 years, 9 days ago)

Quote:

Josex said:
Explain thyself :lol: I did that after stumbling upon one of RR's posts a bit ago when I was researching how to germinate stubborn spores.

Apparently spores can withstand agar at 122f for a short time. If they germinate they'll do it under the layer of poured agar and will eventually resurface leaving any possible bacteria behind.




As I like to source any findings, with me being a noob and all, do you think you might be able to get me a link to his thread that you speak of.


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InvisibleTookitooki
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Re: Let's All Grow Mushrooms 2017 [Re: Tookitooki]
    #24028086 - 01/21/17 12:10 PM (7 years, 9 days ago)

Bod, how do you measure the temp of your agar.  I've been trying to minimize my condensation in my plate.  I give my PC about 1.5-2 hours to cool, open the lid and take watch the temp of the water in the PC until it gets down to about 125 then pull it out.  It take me about 5 mins from that point until I start pouring and I still get condensation in about half of them.


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Re: Let's All Grow Mushrooms 2017 [Re: Tookitooki]
    #24028309 - 01/21/17 02:07 PM (7 years, 9 days ago)

Quote:

Tookitooki said:
Bod, how do you measure the temp of your agar.  I've been trying to minimize my condensation in my plate.  I give my PC about 1.5-2 hours to cool, open the lid and take watch the temp of the water in the PC until it gets down to about 125 then pull it out.  It take me about 5 mins from that point until I start pouring and I still get condensation in about half of them.



in my tek I explain the water bath and how that works.
also you can use a infrared thermometer


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InvisibleComebackKid
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Re: Let's All Grow Mushrooms 2017 [Re: Tookitooki]
    #24028362 - 01/21/17 02:38 PM (7 years, 9 days ago)

Damn. All this time I thought higher nutes were good for germination


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Re: Let's All Grow Mushrooms 2017 [Re: Tookitooki]
    #24028385 - 01/21/17 02:51 PM (7 years, 9 days ago)

Quote:

Tookitooki said:
They require a moisture content to permeate it's membrane, to get to intake any kind of substance





i wouldnt say it REQUIRES moisture. ive dropped straight spores on agar. germ in 8days.
was AA+ i got from a shroomy person :wink:

longest spores were random cubes in a syringe. took 13days.


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OfflineTiamo
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Re: Let's All Grow Mushrooms 2017 [Re: ComebackKid]
    #24028387 - 01/21/17 02:51 PM (7 years, 9 days ago)

Me too. I'm sceptical. I think the nutrients don't really matter too much compared to the agar. Softer agar does wonders for spores. I tried to do some research on this site. RR says he uses less nutes to make the agar more "soupy". Don't really know what to make of that.


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OfflineKenetic
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Re: Let's All Grow Mushrooms 2017 [Re: Tookitooki]
    #24028395 - 01/21/17 02:54 PM (7 years, 9 days ago)

I use an infrared thermomer


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InvisibleTookitooki
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Re: Let's All Grow Mushrooms 2017 [Re: mushboy]
    #24028406 - 01/21/17 03:00 PM (7 years, 9 days ago)

I tried the water bath, the water came down to temp and I still got a lot of condensation.  My flask was still hot to the touch.  The next time I prepped agar, I used the PC( was still decently full of water) as my way bath. It helped but still had condensation. Next time I'm going to give it a few more degrees drop before I take it out.  I'm getting faster at pouring my plates so I should be able to knock out a sleeve before it starts to solidify.  My first pour, I didn't get it all in time. I was really slow at it.  I think I'll invest in a infrared thermometer.

Quote:

mushboy said:
Quote:

Tookitooki said:
They require a moisture content to permeate it's membrane, to get to intake any kind of substance





i wouldnt say it REQUIRES moisture. ive dropped straight spores on agar. germ in 8days.
was AA+ i got from a shroomy person :wink:

longest spores were random cubes in a syringe. took 13days.



I'm sure they get moisture from the agar, just miniscule.

If you put malt extract powder on a spore print, would it germinate?


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Re: Let's All Grow Mushrooms 2017 [Re: Tookitooki]
    #24028414 - 01/21/17 03:04 PM (7 years, 9 days ago)

i don't have any. :shrug:

if the dry spores get a 'miniscule' amount from the agar then making it less stiff would be pointless.

im not saying im right or your wrong. just askin yo.

Quote:

Tookitooki said:
The same reason we don't use 90% ISO




i thought we use 70% because 90 evaporates too quickly.


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Offlinefoodguy
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Re: Let's All Grow Mushrooms 2017 [Re: mushboy]
    #24028443 - 01/21/17 03:18 PM (7 years, 9 days ago)

I've got some GT spores on agar a few days ago, also had a decent amount of condensation but not too bad. I just dropped a small drop in the center. Looks like a lot of bacteria so far, with a few spots of white popping through. I think I may try to streak some spores on another couple plates.


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Re: Let's All Grow Mushrooms 2017 [Re: mushboy]
    #24028623 - 01/21/17 04:35 PM (7 years, 9 days ago)

Quote:

Tookitooki said:
The same reason we don't use 90% ISO, is the same reason we might try a LC if they don't germinate on normal agar.



Quote:

mushboy said:


if the dry spores get a 'miniscule' amount from the agar then making it less stiff would be pointless.

Quote:

Tookitooki said:
The same reason we don't use 90% ISO




i thought we use 70% because 90 evaporates too quickly.




Correct, the water content of the 70% is the reason we use it.  It evaps slower allowing it to penetrate the spores. 

(Edited to correct above info)

Quote:

bodhisatta said:
Evaporation certainty helps it's not the reason it works better though. 70 works at a fundamental level differently than 90+%
College professors have been known to be dumbasses...

Higher alcohol concentrations, like 91%-99%, coagulates protein on contact, creating a "shell" around the entire bacterial organism. After that shell is made, the alcohol CANNOT go through that shell to further coagulate the rest of the bacterial organism and its organelles. This will render the bacteria, not dead, but inactive. If given favorable conditions, it can start to function once more. Now if you use 70% alcohol (which has the highest osmotic pressure of alcohol/water mixtures), it will coagulate the protein but it will do so at a much slower rate, giving it the opportunity to penetrate fully. Since it does this slowly, nothing can coagulate in time to stop it which ends up leaving the bacterial organism dead.






I believe softer agar would be a wetter agar thus transferring more moister to the spores.  Harder agar is a dryer agar.  Stubborn spores might not contract enough moisture on hard agar compared to a softer soupier agar.  You also hear of spore syringes germinating and you don't hear of spore prints germinating


Edited by Tookitooki (01/22/17 01:27 PM)


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Re: Let's All Grow Mushrooms 2017 [Re: Tookitooki]
    #24028625 - 01/21/17 04:38 PM (7 years, 9 days ago)

Quote:

Tookitooki said:
It evaps slower allowing it to penetrate the spores (..) You also hear of spore syringes germinating and you don't hear of spore prints germinating




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