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OfflineForevergreen
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Registered: 06/23/20
Posts: 173
Last seen: 3 years, 5 months
2nd time with agar and im aiming for success
    #26788497 - 06/27/20 11:17 PM (3 years, 6 months ago)

Last time, my first time, I fucked up 80 dishes.

I just made my 2nd agar.
15gr agar
12gr MLE
750 ml water (tequila bottles with holes drilled and filter discs inside lid)
X2

I added to water then boiled. As soon as boiled I poured into bottles. Already looks better than last time. When the bottles come out the pressure cooker, they were 200 and 205 degrees F for anyone wanting to know.

Pressure cooked 15-17psi for 30 min. I then filled up my 25x all american autoclave with water and will use as 120degree F water bath.

Once agar is cooled to 120 i will pull one out, wipe with iOS before introducing to SAB and start pouring my dishes once it cools to 100 degrees F Will post pics later.

Last time I had too much condensation from pouring too hot and not allowing to cool before inoculation. I thought it was fine, but I didnt use my temp gun. I also thought it would be fine to just squirt MS liquid into the dishes. Instead of just a drop. Watch me correct my mistakes.

My air purifer is running 3/4 speed (full speed makes too much noise)
I already wiped down everything with lysol. Im going to turn fan off, put dog outside and wipe everything down AGAIN. after which i will wipe down my body with iso, put on mask etc and go to work. See y'all soon.


BTW i made 80 dishes and wrapped with parafilm. All inoculated with spore syringes. Too much moisture led to me throwing them all away. I found three dishes i didnt wrap with parafilm. They all had mold but TWO ACTUALLY HAFAD MYCELIUM. They werent wrapped and just left aside. Proves my theory right that I fucked up and all 80 plates had too much moisture from pouring too hot and not allowing to cool. I could have 80 dishes of mycelium to work with right now if it wasnt for that dumb mistake. I did toss the two plates with mycelium just because they werent even labeled and want to start fresh.


Edited by Forevergreen (06/27/20 11:22 PM)


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OfflineForevergreen
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Registered: 06/23/20
Posts: 173
Last seen: 3 years, 5 months
Re: 2nd time with agar and im aiming for success [Re: Forevergreen]
    #26788639 - 06/28/20 01:17 AM (3 years, 6 months ago)



Here's my agar. In stacks with hot water on top to reduce condensation. Also I haven't wrapped with parafilm and will inoculate in the morning, once I know they are set. And i will only put a drop per plate.


Waterbath for agar. Its my autoclave with water.


Frosty sticky icky


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OfflineTrichodingus
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Registered: 05/31/20
Posts: 85
Loc: Communing with the divine
Last seen: 3 years, 6 months
Re: 2nd time with agar and im aiming for success [Re: Forevergreen]
    #26788652 - 06/28/20 01:26 AM (3 years, 6 months ago)

Have you looked into no-pour agar?

Also, why start with so many plates?
If you do 10 plates with solid technique you could expand them into an absurd amount of spawn.

Not only that, but more plates are harder to manage especially if you have to transfer a few times.

Agar is usually used to isolate clean cultures and specific genetics. It's better to have fewer quality plates and plan on using g2g for getting volume.


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OfflineForevergreen
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Registered: 06/23/20
Posts: 173
Last seen: 3 years, 5 months
Re: 2nd time with agar and im aiming for success [Re: Trichodingus]
    #26788685 - 06/28/20 01:47 AM (3 years, 6 months ago)

Im not using all plates at once. 10 x 4 syringes. Amazon, hawaiian, ecuador, and red boys. The rest are going in the fridge for the transfers after getting wrapped with parafilm. Let me know if im fucking up. Why not do 10 and transfer best isolates. Instead of jumping into MS to grain I'll do agar. Im gonna have 80 qt jars going. 60 needed 20 for contams.

Everyone keeps asking me why so many. Why not. I don't get the hate on jumping into bulk bulk. Ive heard it all and I guess im just ignorant lol so stay along for the ride or get splashed.


Edited by Forevergreen (06/28/20 01:53 AM)


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InvisibleSunny Skies
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Registered: 05/03/17
Posts: 421
Loc: my house
Re: 2nd time with agar and im aiming for success [Re: Forevergreen]
    #26788978 - 06/28/20 06:26 AM (3 years, 6 months ago)

Your cool man..when I PC I try and fit as much in as possible, so duneenworrywifit and do your thing poppa!


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InvisiblePastywhyteMDiscord
Say hello to my little friend
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Registered: 09/15/12
Posts: 37,808
Loc: Canada Flag
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Re: 2nd time with agar and im aiming for success [Re: Sunny Skies]
    #26789086 - 06/28/20 07:46 AM (3 years, 6 months ago)

When doing spores to grain I’ll inoculate at least 4 plates and usually more like 8 with spores. Transfers will only be taken from the best plate or two. From there only the best transfers make the cut. If you start with more spore inoculated plates it gives you more choices to choose from.


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OfflineForevergreen
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Registered: 06/23/20
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Last seen: 3 years, 5 months
Re: 2nd time with agar and im aiming for success [Re: Pastywhyte]
    #26789453 - 06/28/20 10:22 AM (3 years, 6 months ago)

Quote:

Pastywhyte said:
When doing spores to grain I’ll inoculate at least 4 plates and usually more like 8 with spores. Transfers will only be taken from the best plate or two. From there only the best transfers make the cut. If you start with more spore inoculated plates it gives you more choices to choose from.




Thanks pasty!


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OfflineForevergreen
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Registered: 06/23/20
Posts: 173
Last seen: 3 years, 5 months
Re: 2nd time with agar and im aiming for success [Re: Forevergreen]
    #26789456 - 06/28/20 10:23 AM (3 years, 6 months ago)

All the plates have condensation in them so im going to just them all sit without parafilm until most is gone, then i will inoculate and wrap with parafilm.


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OfflineForevergreen
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Registered: 06/23/20
Posts: 173
Last seen: 3 years, 5 months
Re: 2nd time with agar and im aiming for success [Re: Forevergreen]
    #26790988 - 06/28/20 09:34 PM (3 years, 6 months ago)

Its been about 24 hours and after raising room temp to 80 all the plates have no condensation and then agar is perfect. I sacrificed a plate that got exposed to air anyways. Perfect little jelly cake. Will inoculate tonight I just got more iso 70%

I got wbs from Fred meyers and it has a bunch of cracked corn. I just read the ingredients and it didn't say corn. Once I got home I noticed its just littered with cracked corn. I could return it but I dont save my receipts and will just buy a bird feeder. Tractor supply next to me sells oats but I hear people have lost weight just using oats, they go back to wbs and have better flushes.



Edited by Forevergreen (06/28/20 09:41 PM)


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OfflineForevergreen
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Registered: 06/23/20
Posts: 173
Last seen: 3 years, 5 months
Re: 2nd time with agar and im aiming for success [Re: Forevergreen]
    #26797972 - 07/01/20 10:17 PM (3 years, 6 months ago)

I inoculated on 6/29. Checked plates today and none have mycelium, but at least 15 have contams. Will post pics later.

I have WBS soaking. 10 cups with 1/4 cup gypsum. Will pc tomorrow. I have a 50lb bag of oats too.
Im going to inoculate more plates with the syringes tomorrow.


Edited by Forevergreen (07/01/20 10:18 PM)


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OfflineMH5109
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Registered: 09/27/17
Posts: 1,385
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Re: 2nd time with agar and im aiming for success [Re: Forevergreen]
    #26798029 - 07/01/20 10:48 PM (3 years, 6 months ago)

Agar takes practice, the more you practice the better you will become. Did you inoculate with spores or was it from another plate. Cuz spores take a little bit to germinate.


--------------------




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OfflineForevergreen
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Registered: 06/23/20
Posts: 173
Last seen: 3 years, 5 months
Re: 2nd time with agar and im aiming for success [Re: MH5109]
    #26798032 - 07/01/20 10:49 PM (3 years, 6 months ago)

Spores. Theres a bunch of plates with nothing at all so not only are they clean but hopefully the spores takeoff in a week or so. Just sucks to see some white spots and get excited until they turn green the next day.


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