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NightPuma1
Stranger

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


Registered: 01/23/13
Posts: 15,577
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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) |
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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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enlightenment
alchemist


Registered: 08/09/09
Posts: 1,647
Loc: Europe
Last seen: 8 months, 22 days
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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) |
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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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Kush_Zombie
smug piece of shit



Registered: 10/22/14
Posts: 4,793
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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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mushboy
modboy



Registered: 04/24/05
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) |
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spores on agar from a syringe took 12 days to germ one time. wait a bit longer.
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enlightenment
alchemist


Registered: 08/09/09
Posts: 1,647
Loc: Europe
Last seen: 8 months, 22 days
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Re: Failing at Agar, what's up? [Re: Kush_Zombie]
#23979102 - 01/03/17 03:11 PM (7 years, 27 days ago) |
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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.

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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fatmatic
Stranger


Registered: 12/23/16
Posts: 46
Last seen: 6 years, 8 months
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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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Kush_Zombie
smug piece of shit



Registered: 10/22/14
Posts: 4,793
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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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mushboy
modboy



Registered: 04/24/05
Posts: 32,281
Loc: where?
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Re: Failing at Agar, what's up? [Re: Kush_Zombie]
#23979393 - 01/03/17 05:07 PM (7 years, 27 days ago) |
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why waste $20 when you already have spores?
id rather get a pizza and beer
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Kush_Zombie
smug piece of shit



Registered: 10/22/14
Posts: 4,793
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Re: Failing at Agar, what's up? [Re: mushboy] 1
#23979399 - 01/03/17 05:10 PM (7 years, 27 days ago) |
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If you consider that $20 wasted in a period of 5+ years then you probably shouldn't be buying pizza and beer anyways
-------------------- 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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mushboy
modboy



Registered: 04/24/05
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) |
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Adden

Registered: 06/04/03
Posts: 39,201
Loc:
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Re: Failing at Agar, what's up? [Re: NightPuma1]
#23979406 - 01/03/17 05:13 PM (7 years, 27 days ago) |
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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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Kush_Zombie
smug piece of shit



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

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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mushboy
modboy



Registered: 04/24/05
Posts: 32,281
Loc: where?
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Re: Failing at Agar, what's up? [Re: Kush_Zombie]
#23979415 - 01/03/17 05:17 PM (7 years, 27 days ago) |
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jesus, calm down.
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Kush_Zombie
smug piece of shit



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


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



Registered: 07/26/15
Posts: 1,201
Loc: Colonial alley
Last seen: 14 hours, 25 minutes
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Re: Failing at Agar, what's up? [Re: Munchauzen]
#23979511 - 01/03/17 05:45 PM (7 years, 27 days ago) |
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It could take 15 days op.
-------------------- 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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NightPuma1
Stranger

Registered: 03/28/16
Posts: 310
Last seen: 2 years, 10 days
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Re: Failing at Agar, what's up? [Re: Munchauzen]
#23979611 - 01/03/17 06:20 PM (7 years, 27 days ago) |
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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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lovelaughlibs
Dopamine Slave



Registered: 10/14/15
Posts: 1,811
Loc: England
Last seen: 5 years, 11 months
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Re: Failing at Agar, what's up? [Re: NightPuma1]
#23979717 - 01/03/17 07:17 PM (7 years, 27 days ago) |
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Which tek told you to drill your petris?!
-------------------- Ask and ye shall receive; Seek and ye shall find.
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Adden

Registered: 06/04/03
Posts: 39,201
Loc:
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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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