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Amazon Shop: ½ Pint Jars, Agar, Malt Extract, Petri Dish, Pressure Cooker

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OfflineAiko Aiko
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Grain to Agar?
    #4596325 - 08/29/05 09:36 PM (11 years, 9 months ago)

I hope this is'nt a stupid question, but you can start an agar plate whith a small amoount of colonized grain, correct?


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Man says, "God, show me and I will believe." God says, "Believe and I will show you."


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InvisibleTien
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Re: Grain to Agar? [Re: Aiko Aiko]
    #4596334 - 08/29/05 09:39 PM (11 years, 9 months ago)

Yes...and chances are it will be a really nice rhizo isolate.

Pluto


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OfflineNashbar
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Re: Grain to Agar? [Re: Aiko Aiko]
    #4596336 - 08/29/05 09:39 PM (11 years, 9 months ago)

yup, works fine... might need some peroxide or multiple transfers to get a good, isolated plate. It's definitely not difficult, just takes a couple weeks.


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OfflineAiko Aiko
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Re: Grain to Agar? [Re: Nashbar]
    #4596407 - 08/29/05 10:01 PM (11 years, 9 months ago)

Awesome, that's exactly what I wanted yo hear. BTW, do you guys mix your own agar, or do you buy it premade?


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OfflineNashbar
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Re: Grain to Agar? [Re: Aiko Aiko]
    #4596445 - 08/29/05 10:14 PM (11 years, 9 months ago)

homemade

if you're interested in real sterility, 1/2 and 1/4 pint jars work real well to pressure cook your agar.

agar agar + light malt dextrose + water

heat until melted and dissolved, let cool to 130F, then add peroxide. Don't sterilize the peroxide agar, it will decay to water and oxygen above ~140F


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:pacman: :mushroom2:



Edited by Nashbar (12/13/06 07:20 PM)


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OfflineAiko Aiko
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Re: Grain to Agar? [Re: Nashbar]
    #4596460 - 08/29/05 10:19 PM (11 years, 9 months ago)

What are your ratio's?


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OfflineNashbar
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Re: Grain to Agar? [Re: Aiko Aiko]
    #4596471 - 08/29/05 10:22 PM (11 years, 9 months ago)

1:1:50

agar agar:dextrose:water

reduce the amount of water according to how much peroxide you're gonna add. There is a good TEK somewhere.


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OfflineNashbar
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Re: Grain to Agar? [Re: Nashbar]
    #4596480 - 08/29/05 10:23 PM (11 years, 9 months ago)

couldn't find the TEK, but I saved it since I'm poor and my internet connection will be the first to go if I can't pay the bills.

First you need to get a hold of some light malt extract powder and some agar agar. Malt extract can be found at your local wine and beer brewing store, and agar agar can be bought from Fungi Perfecti, among other places. You also need some petri dishes, also available from FP, or you can pour your agar into empty jars instead of plates if you prefer. Next, mix three grams of malt extract and three grams of agar (or, if you bought them pre-mixed, six grams of the mix) with 149 ml of water (you should use 149 ml instead of 150 because you want to account for an extra ml of peroxide that you will be adding later). I measure out my water with an empty syringe, keeping in mind that one cc is the same as one ml. I mix and sterilize this solution in a half pint jar, because the amount of agar solution is right for that size jar and because the size of the jar fits well in the pot that I use for sterilizing it. It works for me, but it is not ideal for pouring and I'm sure a better substitute could be found. Anyway, close up your container of agar and sterilize for 45 minutes. I sterilize mine by letting it sit in boiling water in a covered pot, but you could use a pressure cooker if you wanted to. While it is sterilizing, thoroughly clean your plates if you haven't already done so, and stack them on a tabletop. As soon as is possible without burning yourself, move the jar of molten agar to the tabletop, open the lid, and insert a clean thermometer into the agar. Sine hydrogen peroxide decomposes at 140 degrees fahrenheit, you need to let the temperature of the agar drop below that before you add the peroxide. Otherwise it will decompose into water and oxygen and be effectively useless. This is why the peroxide is added after sterilization. I have found that my agar begins to solidify at around 120 degrees fahrenheit, so fill a syringe with peroxide and when the temperature falls to 130 degrees, add one ml to the molten agar and stir it in with the thermometer. Then remove the thermometer and pour your plates, stacking them as you go. I can usually pour nine or ten plates with this amount of agar. If you want to make a larger or smaller amount, simply adjust the amounts of water, agar, malt, and peroxide, keeping all the ratios the same.

A good idea would be to order some antibiotic agar from FP to use for germinating spores. This stuff, while resistant to bacteria, is still susceptible to contamination by mold and yeast, so don't forget to use the proper sterile techniques (like a glove box, bleached down room, or a warm oven). Once the spores germinate, the mycelium can be transferred to peroxidated agar with less of a stress on sterility. It is important to remember to keep your scalpels clean and to thoroughly clean and sterilize your plates prior to pouring! If there are small mold colonies left in the plates when you pour them, they will be resistant to the peroxide and will grow.

Commentary added by mycofile 5-5-2000: A range of H2O2 concentrations is acceptable. cubensis seems to tolerate amounts ranging from 1 ml to 7ml for this mix. The more you use, the longer "adjustment" period for mycelium to adapt to the peroxidated agar. As a general rule, the least amount necessary is best. But since your mileage may varry, you may modify as to what works for you.


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OfflineAiko Aiko
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Re: Grain to Agar? [Re: Nashbar]
    #4596516 - 08/29/05 10:31 PM (11 years, 9 months ago)

Nice, that helps alot! Thanx man. I'm new to agar, but it really sounds interesting and efficient. I'll let you know how it turns out. Probably in the next few weeks. Peace! :gd_icon: :beer: :mushroomgrow:


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Man says, "God, show me and I will believe." God says, "Believe and I will show you."


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OfflineNashbar
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Re: Grain to Agar? [Re: Aiko Aiko]
    #4596528 - 08/29/05 10:34 PM (11 years, 9 months ago)

it's kinda fun too, isolated a few strains from random bags bought on the street. Cheap, efficient and entertaining, can't ask for much more.


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Edited by Nashbar (12/13/06 07:18 PM)


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Re: Grain to Agar? [Re: Aiko Aiko]
    #23261248 - 05/23/16 12:20 PM (1 year, 1 month ago)

You can but you might as wait to you have a nice fast, fat growing or the charactaristics you desire and then clone that one.


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OfflineJosex
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Re: Grain to Agar? [Re: Mush-e]
    #23261360 - 05/23/16 12:55 PM (1 year, 1 month ago)

Quote:

Mush-e said:
You can but you might as wait to you have a nice fast, fat growing or the charactaristics you desire and then clone that one.




Man you keep resurrecting threads from the graveyard. There's a reason a warning pops up before you bump an old thread; the peeps who posted here are no longer around and even if they were, they got the answers they wanted at the time.

Plus, old threads are normally full of bad and outdated info, like this one for example.

Plus, plus, you give terrible advice :facepalm:


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OnlinebodhisattaM
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Re: Grain to Agar? [Re: Mush-e]
    #23261378 - 05/23/16 01:00 PM (1 year, 1 month ago)

Quote:

Mush-e said:
You can but you might as wait to you have a nice fast, fat growing or the charactaristics you desire and then clone that one.



Are you down syndrome dude?

Your advice is 100% shit and you are posting in 10+ year old threads


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Amazon Shop: ½ Pint Jars, Agar, Malt Extract, Petri Dish, Pressure Cooker

Mushrooms, Mycology and Psychedelics >> Mushroom Cultivation

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