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OfflineNightPuma1
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Registered: 03/28/16
Posts: 310
Last seen: 2 years, 10 days
Failing at Agar, what's up?
    #23979059 - 01/03/17 02:46 PM (7 years, 27 days ago)

Did a no pour tek with metal petri-dishes with glass lids using this recipe:

½ cup of water
2g of Agar
3g of potato flakes (Idahoan original instant mashed potatoes)
1g of honey

This made 10 petri dishes. I pressure cooked them for 40 minutes, inoculated them with a drop from a MS syringe, waited about 5 days at 77F and there is no growth. Not including pictures because there is pretty much nothing to show.

Any thoughts?


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OfflineMorel Guy
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Re: Failing at Agar, what's up? [Re: NightPuma1] * 1
    #23979066 - 01/03/17 02:50 PM (7 years, 27 days ago)

Was the agar cool and firm?


--------------------
"in sterquiliniis invenitur in stercore invenitur"

In filth it will be found in dung it will be found


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Offlineenlightenment
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Posts: 1,647
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Re: Failing at Agar, what's up? [Re: Morel Guy] * 1
    #23979070 - 01/03/17 02:55 PM (7 years, 27 days ago)

It could take more than 5 days to see growth sometimes.
Did you flame the needle and pushed out one drop straight to the plate?
If I use a syringe I push out a few drops to cool the needle. Then I inoculate the agar.


Edited by enlightenment (01/03/17 03:00 PM)


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InvisibleKush_Zombie
smug piece of shit
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Registered: 10/22/14
Posts: 4,793
Re: Failing at Agar, what's up? [Re: enlightenment] * 2
    #23979084 - 01/03/17 03:03 PM (7 years, 27 days ago)

I did agar with a syringe once and it completely went to shit. I got some sterile cotton swabs and ordered a spore print after that and never had an issue since then.


--------------------
How to get started in bulk:
Presto 23-Quart Pressure Cooker
BOD's Simple as FUCK Still Air Box
PastyWhyte's Easy Agar Tek
Munchauzen's Cultivation Video Series
How EvilMushroom666 Prepares His Grains (I use jars with Synthetic Filter Discs)
What is G2G? (Grain-to-Grain)
Damion5050's Coir Tek (I use 5.5 - 6 quarts of water instead of 4. Also ignore step 13 and ignore the monotub completely. The only purpose of this tek is to show you how to make a simple substrate. I also add gypsum to it but not necessary)
Spitball's Monotub Tek (A liner isn't necessary but is useful)
Use 6500k lights throughout the whole process. When you wake up, turn the light on. When you go to sleep turn the light off. It's as simple as that.


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InvisiblemushboyMDiscord
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Posts: 32,281
Loc: where?
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Re: Failing at Agar, what's up? [Re: Kush_Zombie]
    #23979098 - 01/03/17 03:08 PM (7 years, 27 days ago)

spores on agar from a syringe took 12 days to germ one time. wait a bit longer.


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Offlineenlightenment
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Registered: 08/09/09
Posts: 1,647
Loc: Europe
Last seen: 8 months, 21 days
Re: Failing at Agar, what's up? [Re: Kush_Zombie]
    #23979102 - 01/03/17 03:11 PM (7 years, 27 days ago)

Quote:

Kush_Zombie said:
I did agar with a syringe once and it completely went to shit. I got some sterile cotton swabs and ordered a spore print after that and never had an issue since then.




:justno:

Maybe you had bad luck and a bad syringe.

Try to use swabs with a 5+year old print. Without hydration the spores have a very hard time to germinate. You need to reduce the agar content or hydrate the spores in a syringe.
Usually I use swabs but I never had a spore syringe that did not germinate if it was used properly.

With young prints swabs and syringes work well.


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Offlinefatmatic
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Registered: 12/23/16
Posts: 46
Last seen: 6 years, 8 months
Re: Failing at Agar, what's up? [Re: enlightenment]
    #23979258 - 01/03/17 04:14 PM (7 years, 27 days ago)

I have a couple agar plates going on 9 days with nothing. waiting sucks but its the name of the game


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InvisibleKush_Zombie
smug piece of shit
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Registered: 10/22/14
Posts: 4,793
Re: Failing at Agar, what's up? [Re: enlightenment]
    #23979387 - 01/03/17 05:04 PM (7 years, 27 days ago)

First off, when did I say it didn't geminate?

Second, who the hell is holding onto a print for 5 years. And even if someone is, god forbid they have to make a $20 investment on another print in a 5 year timespan.


--------------------
How to get started in bulk:
Presto 23-Quart Pressure Cooker
BOD's Simple as FUCK Still Air Box
PastyWhyte's Easy Agar Tek
Munchauzen's Cultivation Video Series
How EvilMushroom666 Prepares His Grains (I use jars with Synthetic Filter Discs)
What is G2G? (Grain-to-Grain)
Damion5050's Coir Tek (I use 5.5 - 6 quarts of water instead of 4. Also ignore step 13 and ignore the monotub completely. The only purpose of this tek is to show you how to make a simple substrate. I also add gypsum to it but not necessary)
Spitball's Monotub Tek (A liner isn't necessary but is useful)
Use 6500k lights throughout the whole process. When you wake up, turn the light on. When you go to sleep turn the light off. It's as simple as that.


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InvisiblemushboyMDiscord
modboy
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Registered: 04/24/05
Posts: 32,281
Loc: where?
Trusted Cultivator
OG Cultivator
Re: Failing at Agar, what's up? [Re: Kush_Zombie]
    #23979393 - 01/03/17 05:07 PM (7 years, 27 days ago)

why waste $20 when you already have spores?

id rather get a pizza and beer


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InvisibleKush_Zombie
smug piece of shit
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Registered: 10/22/14
Posts: 4,793
Re: Failing at Agar, what's up? [Re: mushboy] * 1
    #23979399 - 01/03/17 05:10 PM (7 years, 27 days ago)

If you consider that $20 wasted in a period of 5+ years then you probably shouldn't be buying pizza and beer anyways :shrug:


--------------------
How to get started in bulk:
Presto 23-Quart Pressure Cooker
BOD's Simple as FUCK Still Air Box
PastyWhyte's Easy Agar Tek
Munchauzen's Cultivation Video Series
How EvilMushroom666 Prepares His Grains (I use jars with Synthetic Filter Discs)
What is G2G? (Grain-to-Grain)
Damion5050's Coir Tek (I use 5.5 - 6 quarts of water instead of 4. Also ignore step 13 and ignore the monotub completely. The only purpose of this tek is to show you how to make a simple substrate. I also add gypsum to it but not necessary)
Spitball's Monotub Tek (A liner isn't necessary but is useful)
Use 6500k lights throughout the whole process. When you wake up, turn the light on. When you go to sleep turn the light off. It's as simple as that.


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InvisiblemushboyMDiscord
modboy
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Posts: 32,281
Loc: where?
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Re: Failing at Agar, what's up? [Re: Kush_Zombie]
    #23979403 - 01/03/17 05:13 PM (7 years, 27 days ago)

:uhno:


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InvisibleAdden
I'm a teapot
Registered: 06/04/03
Posts: 39,201
Loc: Flag
Re: Failing at Agar, what's up? [Re: NightPuma1]
    #23979406 - 01/03/17 05:13 PM (7 years, 27 days ago)

Quote:

NightPuma1 said:
Did a no pour tek with metal petri-dishes with glass lids using this recipe:

½ cup of water
2g of Agar
3g of potato flakes (Idahoan original instant mashed potatoes)
1g of honey

This made 10 petri dishes. I pressure cooked them for 40 minutes, inoculated them with a drop from a MS syringe, waited about 5 days at 77F and there is no growth. Not including pictures because there is pretty much nothing to show.

Any thoughts?




I run my PDA:

7g agar
1.5 cups water
3 drops Karo
1tsp potato flakes

If I'm germinating I'll add another quarter cup to the mixture because softer agar (but not too soft) is ideal for cube spores IME.


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InvisibleKush_Zombie
smug piece of shit
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Registered: 10/22/14
Posts: 4,793
Re: Failing at Agar, what's up? [Re: mushboy]
    #23979411 - 01/03/17 05:16 PM (7 years, 27 days ago)

Quote:

mushboy said:
:uhno:



That's all you can really reply at that point.


--------------------
How to get started in bulk:
Presto 23-Quart Pressure Cooker
BOD's Simple as FUCK Still Air Box
PastyWhyte's Easy Agar Tek
Munchauzen's Cultivation Video Series
How EvilMushroom666 Prepares His Grains (I use jars with Synthetic Filter Discs)
What is G2G? (Grain-to-Grain)
Damion5050's Coir Tek (I use 5.5 - 6 quarts of water instead of 4. Also ignore step 13 and ignore the monotub completely. The only purpose of this tek is to show you how to make a simple substrate. I also add gypsum to it but not necessary)
Spitball's Monotub Tek (A liner isn't necessary but is useful)
Use 6500k lights throughout the whole process. When you wake up, turn the light on. When you go to sleep turn the light off. It's as simple as that.


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InvisiblemushboyMDiscord
modboy
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Registered: 04/24/05
Posts: 32,281
Loc: where?
Trusted Cultivator
OG Cultivator
Re: Failing at Agar, what's up? [Re: Kush_Zombie]
    #23979415 - 01/03/17 05:17 PM (7 years, 27 days ago)

jesus, calm down.:thumbdown:

:badmanthisfeels:


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InvisibleKush_Zombie
smug piece of shit
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Registered: 10/22/14
Posts: 4,793
Re: Failing at Agar, what's up? [Re: mushboy]
    #23979418 - 01/03/17 05:18 PM (7 years, 27 days ago)

That's something I'd say if I wasnt calm.


--------------------
How to get started in bulk:
Presto 23-Quart Pressure Cooker
BOD's Simple as FUCK Still Air Box
PastyWhyte's Easy Agar Tek
Munchauzen's Cultivation Video Series
How EvilMushroom666 Prepares His Grains (I use jars with Synthetic Filter Discs)
What is G2G? (Grain-to-Grain)
Damion5050's Coir Tek (I use 5.5 - 6 quarts of water instead of 4. Also ignore step 13 and ignore the monotub completely. The only purpose of this tek is to show you how to make a simple substrate. I also add gypsum to it but not necessary)
Spitball's Monotub Tek (A liner isn't necessary but is useful)
Use 6500k lights throughout the whole process. When you wake up, turn the light on. When you go to sleep turn the light off. It's as simple as that.


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InvisibleMunchauzen
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Registered: 06/22/11
Posts: 14,342
Trusted Cultivator
Re: Failing at Agar, what's up? [Re: NightPuma1]
    #23979493 - 01/03/17 05:39 PM (7 years, 27 days ago)

It sounds like OP is using my agar recipe which is actually the perfect consistency for germinating spores. I'm not sure how well petri's would work in a no-pour scenario thou? I'd love to see a pic of what they look like.

5 days is nothing. have some patience.


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Offlinewtfcrazymofo
foil hater
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Registered: 07/26/15
Posts: 1,201
Loc: Colonial alley
Last seen: 11 hours, 16 minutes
Re: Failing at Agar, what's up? [Re: Munchauzen]
    #23979511 - 01/03/17 05:45 PM (7 years, 27 days ago)

It could take 15 days op.:chillpill:


--------------------
If you want to eat->https://www.shroomery.org/forums/showflat.php/Number/8553541

Bag sealers are to bulky (my hood isn't that big)
https://www.shroomery.org/forums/showflat.php/Number/28622922




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OfflineNightPuma1
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Posts: 310
Last seen: 2 years, 10 days
Re: Failing at Agar, what's up? [Re: Munchauzen]
    #23979611 - 01/03/17 06:20 PM (7 years, 27 days ago)

Ok so I left something out because I was in a bit of a hurry, but I don't think it should matter much. You let me know:

The petri dishes are metal with glass lids. I drilled a hole in the side towards the top because the tek says to put a synthetic filter over the hole. Well I couldn't glue it on right even though I was using high temperature RTV, so I instead stuffed the hole with polyfil. When I did my injections, I just injected through the polyfil like I was inoculating a jar of grain. It seems like a good idea as you get to do both no pour AND I don't have to open the petri dish to inject the solution, thereby reducing contamination.

Thoughts?


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Offlinelovelaughlibs
Dopamine Slave
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Registered: 10/14/15
Posts: 1,811
Loc: England Flag
Last seen: 5 years, 11 months
Re: Failing at Agar, what's up? [Re: NightPuma1]
    #23979717 - 01/03/17 07:17 PM (7 years, 27 days ago)

Which tek told you to drill your petris?!


--------------------
Ask and ye shall receive; Seek and ye shall find.


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InvisibleAdden
I'm a teapot
Registered: 06/04/03
Posts: 39,201
Loc: Flag
Re: Failing at Agar, what's up? [Re: lovelaughlibs]
    #23979758 - 01/03/17 07:32 PM (7 years, 27 days ago)

At this point just buy some pp5's. If you can't, small jars will do. I do my tissue transfers in short glass jars and use big pp5's for spore wipes or myc selection. I don't know how to do slants so I keep cultures in half pint jars leaned up against the PC during sterilization. 1/4" hole with two layers of micropore tape. I can even wash them in the dishwasher six or so times and not run through tape. I use small glad mini rounds for tiger drops into wide mouth quart.


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OfflineKenetic
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Registered: 08/24/14
Posts: 4,389
Loc: I don't believe in land Flag
Last seen: 5 years, 3 months
Re: Failing at Agar, what's up? [Re: Adden]
    #23979765 - 01/03/17 07:34 PM (7 years, 27 days ago)

I've had plenty of plates not germinate for whatever reason. 
Actually is was only with the ms syringes that this happened.  I shook it up really good and put one drop on each dish, 5 dishes total.  Nothing happened.

Later I busted out the syringe again and shook it up real good and put 3 drops on each dish.  5 more dishes.  Bam, a few days later I had some results.
Just do it again.



--------------------
Todo Cambia
   

               
                                                :cookiemonster::elmo:



DMT said: Everyone know's me, they just don't know it yet


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OfflineNightPuma1
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Last seen: 2 years, 10 days
Re: Failing at Agar, what's up? [Re: lovelaughlibs]
    #23980146 - 01/03/17 10:13 PM (7 years, 27 days ago)

OPB Reusable No Pour tek. I made the petris exactly as he does except I use polyfil instead of synthetic filter discs.


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