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Jsneeb
Drifter


Registered: 04/05/15
Posts: 109
Last seen: 6 years, 10 months
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Nothing growing on my agar, Literally nothing, I licked a plate still nothing.
#21806989 - 06/14/15 04:03 PM (8 years, 7 months ago) |
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I have had trouble finding Light Malt Extract locally. Should have just bought some a long time ago thinking I could find it.
My procedure is as follows.
Boil potatoes in water for 30min. Strain potatoes and eat.
Take ~~ 200ml of potato broth and boil it down to 60 ml. ( I have since learned this step is counter intuitive and wrong. I was boiling it down since I thought this would result in more concentrated nutrients, which it does. But this apparently makes for slower growth on the agar as Mycelium doesn't need to reach out as far to search for nutrients. I will not be doing this for the future)
Measure 1.2 grams honey,
Measure 1.2 grams agar powder
Mix everything ( I mix the honey and agar into the still hot potato broth.
Mix until everything dissolves.
Use syringe to inject 10 CC's into 4 oz glass mini jars.
Make 6 of them.
Load into pressure cooker and cook for 45 minutes.
Allow to cool
As I open the PC i quickly snug down all the lids
Lids have a 1/2 inch hole with tight polyfill
No filter disk as I couldn't Find TYVEK at my mail store or Wallmart or the hobby place. ( Found some at Home Depot but it is the Tyvek Home Wrap, would that work? Its thick, like a tarp.)
So I practiced with about 3 plates last week on monday. I took a piece of shroom and cut a slit with my exacto knife in the agar and placed it there.
One plate I left open in my SAB the whole time I worked to use as a control and watch for contams so I would know what they look like.
I posted and found out that I probably doomed this set anyway because I soaked my shroom piece in hydrogen peroxide solution before hand thinking this would clean the sample up.
On friday I did all of this again, used shroom pieces on 3 plates, spores from a print on 3 plates, and spores from syringe on 3 plates. Again I had a control.
Nothing has grown in the monday set but like I mentioned it was probably botched.
Nothing has grown on the friday set either ( only been two days maybe I'll report back)
For fun I licked the control plate from monday which had grown nothing on it during the week.
Its been two days and I dont see anything on it either?
WTF is up with my agar? It has nutrients, why is nothing growing?
-------------------- Read my trip report Titled, Mushrooms, Thirsty Water Bottle, And Boiling Peanut-butter? http://www.reddit.com/r/shrooms/comments/37z9rp/mushrooms_thirsty_water_bottle_and_boiling/
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jboredone
Money-The root of all evil....



Registered: 01/19/12
Posts: 4,783
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Re: Nothing growing on my agar, Literally nothing, I licked a plate still nothing. [Re: Jsneeb]
#21806992 - 06/14/15 04:04 PM (8 years, 7 months ago) |
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you licked it?.....
-------------------- Peace Pot Micro-Dot God Loves You High or Not!!! In order to grow old and wise, you must once have been young and dumb!

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ShroominMe
Stranger
Registered: 05/03/15
Posts: 525
Last seen: 8 years, 3 months
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Re: Nothing growing on my agar, Literally nothing, I licked a plate still nothing. [Re: jboredone]
#21807033 - 06/14/15 04:15 PM (8 years, 7 months ago) |
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I think it took 5 days for me to start seeing myc on my premade agar plates.
Why would you lick your agar plate???
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thoraxx
Wizard


Registered: 12/27/13
Posts: 580
Loc: Bavaria
Last seen: 4 years, 10 months
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Re: Nothing growing on my agar, Literally nothing, I licked a plate still nothing. [Re: jboredone]
#21807044 - 06/14/15 04:19 PM (8 years, 7 months ago) |
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peroxide shouldnt kill mycelium, dunking a piece in there before trying to clone is a smart move
It takes a while for plates to show growth tho, especially when you start from spores Give it a week, if literally nothing grows you found a miracle preservative
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,889
Loc: Milky way
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Re: Nothing growing on my agar, Literally nothing, I licked a plate still nothing. [Re: thoraxx]
#21807074 - 06/14/15 04:27 PM (8 years, 7 months ago) |
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Peroxide has no place in agar work except the catalase test. Check out the third link in my signature
Quit over thinking agar and just follow directions till you can do it.
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eatyualive
Eat's You Alive :)



Registered: 08/17/01
Posts: 19,026
Loc: In Your Head
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Re: Nothing growing on my agar, Literally nothing, I licked a plate still nothing. [Re: bodhisatta]
#21807158 - 06/14/15 04:51 PM (8 years, 7 months ago) |
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can you post a picture of the licked plate when it germinates?
otherwise you may have the cleanest mouth in the universe.
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,889
Loc: Milky way
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Re: Nothing growing on my agar, Literally nothing, I licked a plate still nothing. [Re: eatyualive]
#21807168 - 06/14/15 04:53 PM (8 years, 7 months ago) |
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yea that's really bizarre there's no way it's not growing it's not enough sugar to make it too sugar rich to grow well for bacteria.
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Sksoul
Pan handler



Registered: 10/31/14
Posts: 397
Loc: Far East
Last seen: 3 years, 6 months
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Re: Nothing growing on my agar, Literally nothing, I licked a plate still nothing. [Re: Jsneeb]
#21807195 - 06/14/15 05:00 PM (8 years, 7 months ago) |
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This isn't so much solving your problem as it is making it easier for you next time, but don't mix agar into hot water. It clumps up immediately. Mix it into the cold water and then heat.
Try grain water or corn syrup over potato flakes. I hate having grit at the bottom of my plates.
-------------------- Like all great travellers, I have seen more than I remember, and remember more than I have seen.
Edited by Sksoul (06/14/15 05:32 PM)
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,889
Loc: Milky way
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Re: Nothing growing on my agar, Literally nothing, I licked a plate still nothing. [Re: Sksoul]
#21807202 - 06/14/15 05:02 PM (8 years, 7 months ago) |
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boil and strain your home made agar then PC it. boom easy.
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Sksoul
Pan handler



Registered: 10/31/14
Posts: 397
Loc: Far East
Last seen: 3 years, 6 months
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Re: Nothing growing on my agar, Literally nothing, I licked a plate still nothing. [Re: bodhisatta]
#21807283 - 06/14/15 05:21 PM (8 years, 7 months ago) |
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Quote:
bodhisatta said: boil and strain your home made agar then PC it. boom easy.
Why bother? I don't see an advantage to using potato flakes and straining when you have grain water, or can just boil some potatoes for dinner and use clean water for pda.
-------------------- Like all great travellers, I have seen more than I remember, and remember more than I have seen.
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RyeJar
StrangerDanger


Registered: 04/16/15
Posts: 523
Last seen: 9 months, 30 days
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Re: Nothing growing on my agar, Literally nothing, I licked a plate still nothing. [Re: Sksoul]
#21807298 - 06/14/15 05:26 PM (8 years, 7 months ago) |
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Quote:
Sksoul said:
Quote:
bodhisatta said: boil and strain your home made agar then PC it. boom easy.
Why bother? I don't see an advantage to using potato flakes and straining when you have grain water, or can just boil some potatoes for dinner and use clean water for pda.
Not a fan of potato flakes either so far.
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Jsneeb
Drifter


Registered: 04/05/15
Posts: 109
Last seen: 6 years, 10 months
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Re: Nothing growing on my agar, Literally nothing, I licked a plate still nothing. [Re: RyeJar]
#21807561 - 06/14/15 06:39 PM (8 years, 7 months ago) |
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Grain water? You mean like the rusty looking stuff from the simmering phase of prepping jars? I could try that.
I thought the peroxide would be a smart move too but others said it probably killed my mycelium. I only diluted it 1:1 and there was significant bubbles of oxygen coming from the shroom so I assume the cell walls and everything got screwed hard.
Also why did I lick it? To see if something would grow on it? 2 days and nothing? I want to be sure my agar even works. I thought contams would show up fast so I licked it. My WBS jars have growth but the same spore syringe on agar shows nothing?
I sure will post a picture if and when something germinates on it.
-------------------- Read my trip report Titled, Mushrooms, Thirsty Water Bottle, And Boiling Peanut-butter? http://www.reddit.com/r/shrooms/comments/37z9rp/mushrooms_thirsty_water_bottle_and_boiling/
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BkillsU
Stranger



Registered: 05/10/15
Posts: 58
Last seen: 2 years, 8 months
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Re: Nothing growing on my agar, Literally nothing, I licked a plate still nothing. [Re: Jsneeb]
#21807597 - 06/14/15 06:49 PM (8 years, 7 months ago) |
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My problem with agar is that I can't get anything to grow but mold and bacteria. It's obviously a result of my sterile technique but I've been diligent and still nothing but mold every time, tried spores, cloning from pins, I've tried everything, agar just hates my mycelium. Oh well :/
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