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myo6
Stranger

Registered: 06/06/13
Posts: 21
Last seen: 8 years, 9 months
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Spores germinate on agar but they didn't on grain
#19236146 - 12/06/13 11:42 AM (10 years, 2 months ago) |
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Hello everybody!!
I'm starting to worry,cuz 12 days ago I started to streak some spores on agar; i used the same print to make a syringe in order to inoculate (the same day) 10 hard wheat jars...and they didn't showed sign of growth,after 12 days (usually it take 5 days).
The agar plates are colonizing normally !
My grain prep: a 24 hours soak,followed by a 1h30 sterilization in PC.
My grain jars are made with a GE hole plugged with tight polyfill.
Do you have some ideas?
Thank you in advance!!! myo6
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TheMustardTiger



Registered: 08/19/13
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Re: Spores germinate on agar but they didn't on grain [Re: myo6]
#19236160 - 12/06/13 11:44 AM (10 years, 2 months ago) |
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Sterility issue or genetics? I'm no expert but I'm sure this thread will get some traffic.
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bodhisatta 
Smurf real estate agent



Registered: 04/30/13
Posts: 61,890
Loc: Milky way
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Re: Spores germinate on agar but they didn't on grain [Re: TheMustardTiger]
#19236209 - 12/06/13 11:54 AM (10 years, 2 months ago) |
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post a picture of one of your grain jars. sounds like they're too wet
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Dark76


Registered: 07/14/13
Posts: 121
Last seen: 5 years, 9 months
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Re: Spores germinate on agar but they didn't on grain [Re: myo6]
#19236247 - 12/06/13 12:02 PM (10 years, 2 months ago) |
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And just to be complete...
How did you prepare the syringe?
We could jump straight to the grain if you had used the syringe on both the agar and the grain jars. Since you created the syringe, however, the intermediary steps between print and jar could be the culprit.
Perhaps not likely, but worth a look.
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,811
Loc: Canada
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Re: Spores germinate on agar but they didn't on grain [Re: bodhisatta]
#19236336 - 12/06/13 12:27 PM (10 years, 2 months ago) |
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Quote:
bodhisatta said: post a picture of one of your grain jars. sounds like they're too wet
^This is likely. If your certain thats not the case however, I have many times seen germination on agar outpace germination on grains or brf. Agar is the ideal media for germination of pretty much anything, brf and grains not so much. I have had have some jars inoculated with MS take over two weeks to show growth. Meanwhile the plates I had inoculated at the same time had already been transferred once and master knocked up with wedges. Many times those masters will finish well before the one inoculated with ms solution.
This is all just my experience but I advise a little patience. No matter what, I would wager your plates will provide the better culture in the long run.
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myo6
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Registered: 06/06/13
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Re: Spores germinate on agar but they didn't on grain [Re: Dark76]
#19236346 - 12/06/13 12:31 PM (10 years, 2 months ago) |
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Quote:
Dark76 said: And just to be complete...
How did you prepare the syringe?
We could jump straight to the grain if you had used the syringe on both the agar and the grain jars. Since you created the syringe, however, the intermediary steps between print and jar could be the culprit.
My grain jars aren't to wet,the kernels appears to be on the dry side but they are easily broked using a nail.
For the syringe making,I sterilized tap water in a erlenmeyer (neck stuffed with polyfill,with aluminium foil covering it) during 30 minute,I let it stand during 4 hours waiting the water to be cool, and put some spores on it with a sterile scalpel, and I sucked the water with a sterile syringe. After 12hours,I injected 1ml in each jars.
I forgot to say that I also inoc some drops (the same day)on MEA agar ,(MEA:the same recipe I used for the spores which germinated correctly when I streaked them) whith these syringes....these didn't colonized also......
Thank you for help me,and sorry for my bad english..I really have to improve my english skills ^^
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cronicr



Registered: 08/07/11
Posts: 61,436
Loc: Van Isle
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Re: Spores germinate on agar but they didn't on grain [Re: myo6]
#19236665 - 12/06/13 01:46 PM (10 years, 2 months ago) |
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did you shake your jars after you inoculated?
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  It doesn't matter what i think of you...all that matters is clean spawn I'm tired do me a favor
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Dark76


Registered: 07/14/13
Posts: 121
Last seen: 5 years, 9 months
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Re: Spores germinate on agar but they didn't on grain [Re: myo6]
#19236843 - 12/06/13 02:26 PM (10 years, 2 months ago) |
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Quote:
myo6 said: For the syringe making,I sterilized tap water in a erlenmeyer (neck stuffed with polyfill,with aluminium foil covering it) during 30 minute,I let it stand during 4 hours waiting the water to be cool, and put some spores on it with a sterile scalpel, and I sucked the water with a sterile syringe. After 12hours,I injected 1ml in each jars.
I forgot to say that I also inoc some drops (the same day)on MEA agar ,(MEA:the same recipe I used for the spores which germinated correctly when I streaked them) whith these syringes....these didn't colonized also......
Thank you for help me,and sorry for my bad english..I really have to improve my english skills ^^
Your English is better than any foreign language I speak. 
I would tend to believe something went wrong in the syringe making process. Have you made syringes in the past with success?
First, let me say that I am no expert. We know the spores are viable, but when the syringe made with them is used to inoculate both agar and grain, no growth occurs.
Perhaps, the scalpel was too hot when the spores were scraped. 
Posting pictures would help the experts here offer better guidance.
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SpitballJedi
Ancient Astronaut



Registered: 10/13/12
Posts: 8,598
Loc: Nibiru
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Re: Spores germinate on agar but they didn't on grain [Re: Dark76]
#19236862 - 12/06/13 02:30 PM (10 years, 2 months ago) |
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Quote:
Dark76 said: Perhaps, the scalpel was too hot when the spores were scraped. 
Most likely.
-------------------- The Basics A little civility goes a long way The Noob Forum The Hammock Hangers' Forum
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anne halonium
jaguarette


Registered: 05/07/13
Posts: 1,908
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Re: Spores germinate on agar but they didn't on grain [Re: SpitballJedi]
#19236973 - 12/06/13 02:57 PM (10 years, 2 months ago) |
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bacteria.
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36fuckin5
Alchemycologist


Registered: 08/11/03
Posts: 12,081
Loc: Diving into Mystical Territori...
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Re: Spores germinate on agar but they didn't on grain [Re: anne halonium]
#19237008 - 12/06/13 03:04 PM (10 years, 2 months ago) |
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IT wouldn't be bacteria. That would show up on the agar.
I'd guess it's the tap water. It probably had too much chlorine in it. It also sounds like your grain is too dry. You need to soak it, then SIMMER, then dry it. Soaking alone is not enough.
But if you're doing agar, fuck using a syringe. You're taking a step backwards.
-------------------- Redd Foxx said: If you're offended I don't give a shit and don't come see me no more. Pat The Bunny said: A punk rock song won't ever change the world, but I can tell you about a couple that changed me. bodhisatta said: i recommend common sense and figuring it out. These are the TEKs I use. They're all as cheap and easy as possible, just like your mom.
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anne halonium
jaguarette


Registered: 05/07/13
Posts: 1,908
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Re: Spores germinate on agar but they didn't on grain [Re: 36fuckin5]
#19237020 - 12/06/13 03:09 PM (10 years, 2 months ago) |
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Quote:
36fuckin5 said: IT wouldn't be bacteria. That would show up on the agar.
unless the JARS were bacteria swept.
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36fuckin5
Alchemycologist


Registered: 08/11/03
Posts: 12,081
Loc: Diving into Mystical Territori...
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Re: Spores germinate on agar but they didn't on grain [Re: anne halonium]
#19237027 - 12/06/13 03:10 PM (10 years, 2 months ago) |
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Maybe, but then why would the same syringe not germinate on grain or agar, but he mentions no weird growth or smells on either?
I'm still saying he killed the spores somehow.
-------------------- Redd Foxx said: If you're offended I don't give a shit and don't come see me no more. Pat The Bunny said: A punk rock song won't ever change the world, but I can tell you about a couple that changed me. bodhisatta said: i recommend common sense and figuring it out. These are the TEKs I use. They're all as cheap and easy as possible, just like your mom.
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anne halonium
jaguarette


Registered: 05/07/13
Posts: 1,908
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Re: Spores germinate on agar but they didn't on grain [Re: 36fuckin5]
#19237033 - 12/06/13 03:13 PM (10 years, 2 months ago) |
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he said they did on agar.
its possible he killed his spores also in some kinda handling maneuver.
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36fuckin5
Alchemycologist


Registered: 08/11/03
Posts: 12,081
Loc: Diving into Mystical Territori...
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Re: Spores germinate on agar but they didn't on grain [Re: myo6]
#19237042 - 12/06/13 03:16 PM (10 years, 2 months ago) |
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Quote:
myo6 said: I forgot to say that I also inoc some drops (the same day)on MEA agar ,(MEA:the same recipe I used for the spores which germinated correctly when I streaked them) whith these syringes....these didn't colonized also....
The wording is fucked, but it's fairly clear that he used the same syringe to inoculate jars and petris, and neither germinated. He used the same recipe of agar and streaked some plates properly from the print, which did germinate.
I'm still saying the syringe is the problem.
-------------------- Redd Foxx said: If you're offended I don't give a shit and don't come see me no more. Pat The Bunny said: A punk rock song won't ever change the world, but I can tell you about a couple that changed me. bodhisatta said: i recommend common sense and figuring it out. These are the TEKs I use. They're all as cheap and easy as possible, just like your mom.
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cronicr



Registered: 08/07/11
Posts: 61,436
Loc: Van Isle
Last seen: 2 years, 1 month
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Re: Spores germinate on agar but they didn't on grain [Re: 36fuckin5]
#19237055 - 12/06/13 03:18 PM (10 years, 2 months ago) |
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i'd just start more jars and use the plates that did colonize to inoculate w/o a syringe
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  It doesn't matter what i think of you...all that matters is clean spawn I'm tired do me a favor
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SpitballJedi
Ancient Astronaut



Registered: 10/13/12
Posts: 8,598
Loc: Nibiru
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Re: Spores germinate on agar but they didn't on grain [Re: 36fuckin5]
#19237067 - 12/06/13 03:20 PM (10 years, 2 months ago) |
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He went from print to agar with success.
He went from print to syringe to agar and grain without success.
He only mentioned the scalpel for making the syringe.
I would like to hear more about the steps when using the scalpel.
-------------------- The Basics A little civility goes a long way The Noob Forum The Hammock Hangers' Forum
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cronicr



Registered: 08/07/11
Posts: 61,436
Loc: Van Isle
Last seen: 2 years, 1 month
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Re: Spores germinate on agar but they didn't on grain [Re: SpitballJedi]
#19237071 - 12/06/13 03:21 PM (10 years, 2 months ago) |
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i don't even like scalpels for prints, too sharp but i agree on the more info
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  It doesn't matter what i think of you...all that matters is clean spawn I'm tired do me a favor
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anne halonium
jaguarette


Registered: 05/07/13
Posts: 1,908
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Re: Spores germinate on agar but they didn't on grain [Re: cronicr]
#19237093 - 12/06/13 03:28 PM (10 years, 2 months ago) |
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sounds to me like cooling polyfill top jars, picked up bacteria......injection, making it worse.
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36fuckin5
Alchemycologist


Registered: 08/11/03
Posts: 12,081
Loc: Diving into Mystical Territori...
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Re: Spores germinate on agar but they didn't on grain [Re: anne halonium]
#19237184 - 12/06/13 03:54 PM (10 years, 2 months ago) |
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Quote:
anne halonium said: sounds to me like cooling polyfill top jars, picked up bacteria......injection, making it worse.
Then why was there no germination on the agar plate inoculated with the syringe, and no report of slimy grain or weird smells?
-------------------- Redd Foxx said: If you're offended I don't give a shit and don't come see me no more. Pat The Bunny said: A punk rock song won't ever change the world, but I can tell you about a couple that changed me. bodhisatta said: i recommend common sense and figuring it out. These are the TEKs I use. They're all as cheap and easy as possible, just like your mom.
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