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Mikeallojee
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Antibiotic Malt Extract Agar
#11479245 - 11/18/09 03:20 PM (14 years, 3 months ago) |
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I am a noob and am wanting to create my first agar to start a culture. I have a couple questions first: I couldn't find agar agar locally so I bought the antibiotic malt extract agar from fungi perfecti. Now I need to know what to do, I am not sure if I need to use the malt extract like I would use the agar agar and put honey and the other ingredients in it that I read off the easy liquid culture thread or can I just use the Antibiotic MEA and mix with h2o and PC it?
Second I don't have a clean spore print I will have to extract spores from a dried specimen that I got a while back what is the best way to do this.
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nachohippie
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Re: Antibiotic Malt Extract Agar [Re: Mikeallojee]
#11479805 - 11/18/09 04:33 PM (14 years, 3 months ago) |
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id like to know as well also i found nutrient broth test tubes would these work??
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Doc_T
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Re: Antibiotic Malt Extract Agar [Re: Mikeallojee]
#11479990 - 11/18/09 04:57 PM (14 years, 3 months ago) |
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The stuff didn't come with a recipe? And none on the site? I'm guessing you just mix it with water, but I don't know.
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stonesun
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Re: Antibiotic Malt Extract Agar [Re: Doc_T]
#11480320 - 11/18/09 05:39 PM (14 years, 3 months ago) |
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From FP:
Antibiotic Malt Extract Agar Our antibiotic media prevents bacterial growth while stimulating the growth of mushroom mycelium. Contains the same powerful formulation as our standard Malt Extract Agar (agar, malt, yeast extract and peptone), but with the addition of an antibiotic. For those who have experienced the disaster bacteria wreak upon the spawn laboratory, this media is the solution. 50 grams are recommended per liter of water.
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badman


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Re: Antibiotic Malt Extract Agar [Re: stonesun]
#11480397 - 11/18/09 05:51 PM (14 years, 3 months ago) |
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Walter
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Re: Antibiotic Malt Extract Agar [Re: Mikeallojee]
#11480414 - 11/18/09 05:53 PM (14 years, 3 months ago) |
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The mix you bought already has everything you need in it. "agar, malt, yeast extract and peptone". Add water and PC.
Quote:
Mikeallojee said: Second I don't have a clean spore print I will have to extract spores from a dried specimen that I got a while back what is the best way to do this.
Search the forums for: spores dry caps. There are a number of posts that seem promising.
Good Luck
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charvo
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Re: Antibiotic Malt Extract Agar [Re: Mikeallojee]
#11480472 - 11/18/09 06:01 PM (14 years, 3 months ago) |
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Thats right the antibiotic agar from stamets is ready to go. If you dont want to make a whole liter you can calculate out how much you need. you alrdy know 50gram to 1 liter. If your planning on using H2O2 you have to add it after pressure cooking, and once the agar has cooled to 125f. The agar should start to solidify at about 120f.
I would just use your dirty print, and isolate clean growth away from the contamination, that's how culture work goes sometimes. so make yourself about 20 plates, and get to it.
good luck
Edited by charvo (11/18/09 06:07 PM)
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Mikeallojee
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Re: Antibiotic Malt Extract Agar [Re: charvo]
#11481675 - 11/18/09 08:39 PM (14 years, 3 months ago) |
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Thanks for the great comments and instructions. I have 18 1/4pint jars, just finished washing them and putting in the holes and silicone. The silicone has to harden and it will probably take over night. I have been taking pics to post and taking detailed notes.
Does the container used to mix the agar matter? I was going to use a large measuring bowl. Does it matter on the cleanliness prior to PC. Or does the PC kill everything? What is the best way to let the agar cool without condensation.
I have all the time in the world and don't mind keeping the agar in the pc until everything cools is this a good idea?
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Mikeallojee
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Re: Antibiotic Malt Extract Agar [Re: Mikeallojee]
#11481757 - 11/18/09 08:47 PM (14 years, 3 months ago) |
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I had been thinking that since I have to use a dirty print that I would inoculate several jars and attempt to fruit in the jars. My hope is that by doing so I will be able to collect a quality spore print and use that in the future. Huge concern is that I will use up all my spores and everything will get contaminated and then I will have nothing and have to buy over the net.
Any thoughts on how big of a specimen sample I need to take off the fruit?
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RogerRabbit
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Re: Antibiotic Malt Extract Agar [Re: Mikeallojee]
#11482029 - 11/18/09 09:22 PM (14 years, 3 months ago) |
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You might benefit from the agar/petri dish page and video clip. Pre-mixed MEA weighs 7 grams per tablespoon, so that would be seven tablespoons per liter of water. Start with cold water and the dry agar mixes in by shaking. With hot water it clumps up, but not with cold. I'd suggest using a whiskey bottle like in the video.
I'd try scraping a few tiny pieces of gill tissue onto several plates. Hopefully there will be some spores that will germinate. RR
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charvo
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Re: Antibiotic Malt Extract Agar [Re: Mikeallojee]
#11482121 - 11/18/09 09:32 PM (14 years, 3 months ago) |
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man you sure got alot of reading ahead of you.
first off you should get a flask to pc your agar in, and if this print you speak of is known to be contamed. i would take it to agar and work with it to clean up.
Dont waste your time trying to make spawn. You will most likely use the whole print and end up with nothing but mold and bacteria.
I would find has much info on agar preparation as you can. read read and read some more.
my answer to condensation is dont worry about it while the agar is cooling in a flask, but once you start to pour it into plates. you will stack the plate one on top of another as you go. so pour and stack them vertically. this will help with the condensation.
I will say this thou google search "cynmar"
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Mikeallojee
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Re: Antibiotic Malt Extract Agar [Re: charvo]
#11483211 - 11/19/09 12:20 AM (14 years, 3 months ago) |
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I watched the video a couple of times and it looks like the way to go but i don't have any dishes. I would have to order them online and same goes for the flask. I live in a little town, hard to not raise suspicions. I went into the pharmacy looking for a syringe and the pharmacist knows me and starts asking all kinds of questions about what I am doing and what I need a needle for etc. I told him that I was taking cultures for mushrooms but he just raised his eye brows and shook his head and told me to go to the feed store.
From some of the techs I read jars work okay they are not as good as dishes particularly because I have to take samples from fruit that I have no idea as to how it was handled or by whom. But since this is my only alternative at this time. I am also planning on taking a clone from a shiitake my friend has on his log.
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Mikeallojee
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Re: Antibiotic Malt Extract Agar [Re: RogerRabbit]
#11483229 - 11/19/09 12:28 AM (14 years, 3 months ago) |
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Quote:
RogerRabbit said: You might benefit from the agar/petri dish page and video clip. FP MEA weighs 7 grams per tablespoon, so that would be seven tablespoons per liter of water. Start with cold water and the dry agar mixes in by shaking. With hot water it clumps up, but not with cold. I'd suggest using a whiskey bottle like in the video.
I'd try scraping a few tiny pieces of gill tissue onto several plates. Hopefully there will be some spores that will germinate. RR
I was thinking of working with one gill at a time in a dead air box. I plan on wearing gloves and spraying lysol and alcohol for extra protection. Will it matter if part of the actual gill gets on the agar? Any other precautions I should take.
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Mikeallojee
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Re: Antibiotic Malt Extract Agar [Re: Doc_T]
#11483235 - 11/19/09 12:31 AM (14 years, 3 months ago) |
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Quote:
Doc_T said: The stuff didn't come with a recipe? And none on the site? I'm guessing you just mix it with water, but I don't know.
The recipe said seven gram to one leter of water. two table spoons equal one gram.
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cyb3rtr0n
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Re: Antibiotic Malt Extract Agar [Re: Mikeallojee]
#11483332 - 11/19/09 01:10 AM (14 years, 3 months ago) |
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A pint beer bottle works good as an agar container. I use 500ml water and 25grams of enriched malt extract agar. That is enough to pour 20 100mm petris..
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NoHum
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Re: Antibiotic Malt Extract Agar [Re: cyb3rtr0n]
#11483337 - 11/19/09 01:12 AM (14 years, 3 months ago) |
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***Post deleted***
***Illegal content removed*** -Roadkill
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Edited by Roadkill (11/19/09 10:12 AM)
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Mikeallojee
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Re: Antibiotic Malt Extract Agar [Re: cyb3rtr0n]
#11484893 - 11/19/09 11:28 AM (14 years, 3 months ago) |
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Cyb3 you make a lot of specimen dishes. You must do it large scale. Do you ever use the dishes for liquid cultures?
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stonesun
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Re: Antibiotic Malt Extract Agar [Re: cyb3rtr0n]
#11484946 - 11/19/09 11:37 AM (14 years, 3 months ago) |
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Quote:
cyb3rtr0n said: A pint beer bottle works good as an agar container. I use 500ml water and 25grams of enriched malt extract agar. That is enough to pour 20 100mm petris..

So you use TWO bottles then?
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stonesun
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Re: Antibiotic Malt Extract Agar [Re: Mikeallojee]
#11484961 - 11/19/09 11:39 AM (14 years, 3 months ago) |
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Quote:
Mikeallojee said: Cyb3 you make a lot of specimen dishes. You must do it large scale. Do you ever use the dishes for liquid cultures?
You mean if he inoculates the dishes with LC?
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cyb3rtr0n
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Re: Antibiotic Malt Extract Agar [Re: Mikeallojee]
#11485032 - 11/19/09 11:58 AM (14 years, 3 months ago) |
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No, I just started working with agar. I don't do it large scale, but the dishes can add up when trying to isolate strains. It's interesting, that's why I want to experiment with it.
The sterile petris come in 20-25 pack sleeves. I just went ahead and poured the whole sleeve. The first sleeve of petris I did, I poured too much in each dish, and only ended up making about 12 with the 500ml mix of malt extract agar+water. But, the second sleeve I did, I poured less in each dish, and ended up filling the whole sleeve.
Then I just store the unused petris back in the sleeve and save some of them for when its time to make transfers.
Quote:
Mikeallojee said: Do you ever use the dishes for liquid cultures?
Are you talking about inoculating a liquid culture with an agar/mycelium wedge? You can do that if you want, I haven't yet..
Some people don't use agar to isolate, and grow out mycelium on it from spores or tissue sample just to be sure of no contaminates, then make a LC syring from that.
Edited by cyb3rtr0n (11/19/09 12:16 PM)
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