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B H O
Registered: 03/17/13
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Re: BOD's Comprehensive Agar TEK [Re: A.k.a]
#27394251 - 07/19/21 07:12 PM (2 years, 8 months ago) |
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Serious question here!
So I understand that the temperature of water can’t exceed 212F on its own and that’s why we use a pressure cooker.
A few questions I have is “When the pressure cooker reaches the required pressure, does all the water in it turn to steam?
What if it is filled half way full with water? Does only a certain amount of steam get created and the rest of the water remains?
If only part of the water turns to steam and the rest remains liquid, does the water in its liquid state exceed over 212F?”
If half the pressure cooker is full of water in its liquid state and only the steam is able to reach sterilizing temperature, wouldn’t that mean that our agar hasn’t been completely sterile?
Idk, this is how I prepare my agar and I rarely run into problems but it crossed my mind as I was loading my PC up to the water line of my media bottles.
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bodhisatta
Smurf real estate agent
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Re: BOD's Comprehensive Agar TEK [Re: B H O]
#27394254 - 07/19/21 07:15 PM (2 years, 8 months ago) |
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No just like when water boils at normal pressure not all of it turns to steam.
The water and steam are both over 212 in a pressure cooker.
The energy it takes to overcome the hydrogen bonds in liquid water are a big part of why despite steam being the same temperature as the water it transmits far more energy and thus is the sterilization agent in the cooker.
As long as half of your container is in the steam you're fine also the lid needs a path for steam (and air to escape) whether its loose or filtered
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B H O
Registered: 03/17/13
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Re: BOD's Comprehensive Agar TEK [Re: bodhisatta]
#27394300 - 07/19/21 07:50 PM (2 years, 8 months ago) |
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Yay thanks for taking the time to explain this for me! 🙏
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bodhisatta
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Re: BOD's Comprehensive Agar TEK [Re: B H O]
#27394366 - 07/19/21 08:33 PM (2 years, 8 months ago) |
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Good questions
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Hurriedhelp
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Re: BOD's Comprehensive Agar TEK [Re: bodhisatta]
#27581077 - 12/14/21 11:26 AM (2 years, 3 months ago) |
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Thread necro, but I wanted to thank you for posting this amazing tutorial. It’s immensely helpful!
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Cranial Feature
Familiar
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Re: BOD's Comprehensive Agar TEK [Re: Hurriedhelp]
#27586509 - 12/18/21 06:52 PM (2 years, 3 months ago) |
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This thread def deserves a bump - a practical course in agar, accessible even to me.
Bodi earns all the praise
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Tony6Strings
student
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Re: BOD's Comprehensive Agar TEK [Re: bodhisatta]
#27609783 - 01/06/22 10:04 PM (2 years, 2 months ago) |
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Made my first "plates" today. Used 5 grams brf, 5 grams agar, squeeze of food coloring (next time a couple drops) sterilized in steam pot because I don't have a PC. Poured them in this SAB I made with a hot coffee can. Thank you for these writeups they are great.
-------------------- "We have arrived at truth, and now we find truth is a mystery- a play of joy, creation, and energy. This is source. This is the mystic touchstone that heals and renews. This is the beginning again. This is entheogenic." -Nicholas Sand A box of grain will ease the pain and love will see you through...
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spuitzakje
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Re: BOD's Comprehensive Agar TEK [Re: bodhisatta]
#27612402 - 01/09/22 03:15 AM (2 years, 2 months ago) |
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Quote:
bodhisatta said: the dishes can be stored for years but it's best to make transfers to new dishes at least once a year, take a small rice sized grain from the master plate and put it to a new plate let it colonize the plate nearly completely then put it to refrigeration, this way you can keep your clone cultures or isolates for decades without worrying about senescence occurring.
Why nearly and not completely?
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MysticMycologist
Dirt Sherpa
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Re: BOD's Comprehensive Agar TEK [Re: spuitzakje]
#27612406 - 01/09/22 03:30 AM (2 years, 2 months ago) |
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You don’t want plates grown all the way to the edge. You make transfers from the leading edges of the mycelium, and the very edge of the plate is a good place for bacteria and other nasties to hide.
-------------------- Two eyes to look, One eye to see. Prying open my third eye
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FredNorris
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Appreciate this write up. I used this tek to make plates for the first time yesterday. Much easier than I thought.
After taking the bottle out of the PC, I put it in a sous vide water bath set to 47C. This kept the temp at 117.2F measured with a meat thermometer. I was working from home and it was in the bath for about 2 hours before I had time to pour. I had no problems pouring and only had minor condensation on a few plates.
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Vamonkeys
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Re: BOD's Comprehensive Agar TEK [Re: bodhisatta]
#27632102 - 01/25/22 12:14 PM (2 years, 2 months ago) |
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Hi, I've been doing a ton of reading on agar work, just started my first plates, and looking foreward to no more spore syringe contamination. Bod's writeups have been great and I'm using them all with pretty good success for a re-noob (the TEK now is way different than it was in college in 1996! So much better!)
Question I have is how often / far down the line with agar transfers should I switch up the agar medium. I've read a few places that changing them up helps, just wasn't sure how often that's recommended. 4th,5th,6th transfer?
I have a good SAB setup so not afraid to pour multiple agars to get the best results possible. Thanks
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bakingator
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Re: BOD's Comprehensive Agar TEK [Re: Vamonkeys]
#27637368 - 01/29/22 11:17 AM (2 years, 1 month ago) |
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Bump. Definite nOOb here, absorbing as much as I can from this thread. Have read about 8 pages but question 1) Why wrap loose capped media jar with foil in PC? OP:"after the pressure drops to 0 you can remove the lid of your PC and tighten down the cap on the bottle and take the foil off." I want to thank BOD for the effort to compile this so long ago, like drinking from a fire hose.
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SeaSalt
Registered: 12/08/21
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Hi, noob here. I PC'd my LME Agar last night and the bottle chipped and the Agar boiled over. I used empty jars so the bottle wouldn't fall over and PC' 20in at 15psi. Was the lid too tight?
Due to the chip in the bottle, I'm going to steam sterilize. 1; How tight should the lid be? 2: Should the bottle be above the water? 3: Lastly, when pouring plates, do you parafilm each plate as it's poured or after you've poured all the plates?
Thank you, I hope I'm posting in the right forum.
-------------------- Avatar by Sol_Exposure
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Goatrider
Rhythm Guitarist
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Re: pour agar TEK [Re: SeaSalt]
#27647879 - 02/06/22 04:00 AM (2 years, 1 month ago) |
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Boiling delay. What kind of cooker do you have, you possibly opened the cooker too early. I usually screw lids tight, then loosen a little turn. What kind of media bottle do you use? Agar media benefits from standing in water, as it cools down slowly like a water bath. After pcing i usually wait about an hour or so to take the bottles out.
I let my poured plates rest in the sab a few days unwrapped. After inoculation they get wrapped (i use grafting tape), also the unused get wrapped for storage and later use.
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CorkRockingham
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Anyone ever run into glass 3 chamber dishes?
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0120
Registered: 12/13/21
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Guys what’s another option to know when it’s on proper temperature? I dont have infrared thermometer and can i do that with regular one? or maybe someone has good method?
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Goatrider
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Loc: Germany
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Re: pour agar TEK [Re: 0120]
#27674955 - 02/26/22 11:17 PM (2 years, 30 days ago) |
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Water bath, or if you can hold the bottle without pain.
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baldur
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Re: BOD's Comprehensive Agar TEK [Re: bodhisatta]
#27703515 - 03/21/22 12:56 PM (2 years, 7 days ago) |
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Today I tried to boil the water before pouring it into the GL45 bottles when blending my agar from malt extract syrup and agar powder.
This worked super well, and all my malt extract and agar just dissolved immediately. I still microwaved the bottles until they boiled, but I'm wondering if I could just have skipped that microwave part and went straight to the PC with the bottles?
If so then this is drastically faster than heating the bottles in the microwave. Note that I was using a pretty good induction stove to heat the water, which resulted in a few liters of water boiling in just a couple of minutes.
And isn't it alright to pour just super hot water into the bottles? Or does the agar need for some reason to be heated gradually?
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baldur
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Re: BOD's Comprehensive Agar TEK [Re: bodhisatta]
#27728566 - 04/10/22 10:05 AM (1 year, 11 months ago) |
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How important is this microwave part?
I'm wondering since it's quite time consuming.
I tried pouring just boiling water into the bottles (straight from a boiling pot), and the mixture seemed to mix very well. No separation in the liquid, and nothing sticking to the bottom when I turned the bottle around.
I've noticed separation when I mix water that is not hot enough.
I microwaved the bottles anyway, but didn't see any difference in the bottles before and after that process.
Soooo, is it necessary?
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baldur
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Re: BOD's Comprehensive Agar TEK [Re: bodhisatta]
#27728569 - 04/10/22 10:09 AM (1 year, 11 months ago) |
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I also in order to shorten the time of this operation, I tried this method:
1. Boil water 2. Mix part of the water in a blender with the agar 3. Pour back into pot and mix
Now this resulted in massive separation when I poured it into bottles, so this method wasn't working well.
Then I tried boiling the agar mixture, but that resulted in burnt agar even before it started to boil.
Now I might be able to boil it if I was stirring it constantly. But I figured that I'd read up and ask around more before ruining more MAE
Anyone have some hints on how to make this process better?
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