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Offlinethemycoman
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Getting started with agar, have a few questions
    #14067755 - 03/04/11 06:49 PM (12 years, 10 months ago)

I am just getting started and I plan to build a GB/SAB like described by todlow here:

http://www.shroomery.org/forums/showflat.php/Number/13552603#13552603

Now I am trying to decide on which type of agar to order.

On mycosupply's website they offer three types: Bacteriological Grade Agar, Potato Dextrose Agar, and Malt Extract Agar.

It seems most people use MEA or mix with PDA.

I will be working primarily with king oyster and shiitake but I am looking to try the paddy straw mushroom and Agaricus blazei Murrill as well.

I was wondering if there was one that would be best for these species.

I also had a question about working with slants since I will be receiving one of shiitake 75 soon.

It seems normally you can just use your scalpel to remove a small piece of agar like shown in RR's videos, but I have also read that sometimes you must remove a small piece of the wood if the mycellium on agar does not take on a new plate.

I was unsure how to safely remove a small piece of wood and was hoping somebody could explain that better to me.

Thanks for all your help in advance.


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InvisibleOzzy
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Re: Getting started with agar, have a few questions [Re: themycoman]
    #14067900 - 03/04/11 07:20 PM (12 years, 10 months ago)

I bought premade agar the first time out.:sad:

I reccomend spending the hour to track down agar at the asian stores in your area and malt from the brew store. Then use Evilmushrooms tec to learn how to make your own.

It turned out to be incredibly simple and looking back I kick myself in the arse for not doing it that way to begin with. If you do that, you don't have to wait a week when you run out/pay shipping/......

good luck with whatever you decide.

I haven't heard of trying to snag some wood aside from the agar? It seems that would be very difficult.


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InvisiblePeterthinks
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Re: Getting started with agar, have a few questions [Re: Ozzy]
    #14068525 - 03/04/11 09:53 PM (12 years, 10 months ago)

Check the link in my sig.

http://www.shroomery.org/forums/showflat.php/Number/13359233#13359233

PYDA  good stuff all I have used from day one and every edible I've tried liked it.
Well except for the white button mushroom you normally see in stores.

http://www.shroomery.org/forums/showflat.php/Number/13103735/fpart/1/vc/1


--------------------
Give a man a fire and he will be warm for the rest of the night.
Set a man on fire and he will be warm for the rest of his life.
NEWB NEWBIE NEWCOMER IGNORANT? QUESTIONS?
Click HERE HERE HERE HERE For detailed instructions with pictures on how to grow mushrooms. There is a lot of info on the Shroomery and this is what you need to know.


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Invisibleteknix
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Re: Getting started with agar, have a few questions [Re: Peterthinks]
    #14068550 - 03/04/11 09:57 PM (12 years, 10 months ago)

I have an extra plate that is getting old and I prob wont use, has some browning in it, but most should still be viable, I'll send you that too.

Generally you use an inoculation loop for slants.


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InvisibleDoc_T
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Re: Getting started with agar, have a few questions [Re: Ozzy]
    #14068567 - 03/04/11 10:01 PM (12 years, 10 months ago)

Quote:

Ozzy said:
I reccomend spending the hour to track down agar at the asian stores in your area and malt from the brew store. Then use Evilmushrooms tec to learn how to make your own.




Agreed 100%. Also check FastFred's Media Cookbook. But malt is all I use.


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InvisiblePeterthinks
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Re: Getting started with agar, have a few questions [Re: Doc_T] * 1
    #14068629 - 03/04/11 10:16 PM (12 years, 10 months ago)



--------------------
Give a man a fire and he will be warm for the rest of the night.
Set a man on fire and he will be warm for the rest of his life.
NEWB NEWBIE NEWCOMER IGNORANT? QUESTIONS?
Click HERE HERE HERE HERE For detailed instructions with pictures on how to grow mushrooms. There is a lot of info on the Shroomery and this is what you need to know.


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Offlinethemycoman
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Re: Getting started with agar, have a few questions [Re: teknix]
    #14070032 - 03/05/11 09:09 AM (12 years, 10 months ago)

Quote:

Ozzy said:
I bought premade agar the first time out.:sad:

I reccomend spending the hour to track down agar at the asian stores in your area and malt from the brew store. Then use Evilmushrooms tec to learn how to make your own.

It turned out to be incredibly simple and looking back I kick myself in the arse for not doing it that way to begin with. If you do that, you don't have to wait a week when you run out/pay shipping/......

good luck with whatever you decide.

I haven't heard of trying to snag some wood aside from the agar? It seems that would be very difficult.




Thanks for the suggestion, I will definitely take the time and make my own sounds much better than waiting for it and paying $40+shipping.

Quote:

teknix said:
I have an extra plate that is getting old and I prob wont use, has some browning in it, but most should still be viable, I'll send you that too.

Generally you use an inoculation loop for slants.




Thank you again for helping me out with she shiitake 75, I am trying to do this all OTC without having to order scalpels or inoculation loops is there anything I can get locally to use in place.

I was thinking I could use tweezers to get a piece out of the slant and an exacto blade (sterilized in foil of course) until I get the proper tools.

Also since I don't have disposable dishes I was thinking of using the short wide mouth jars like Doc_T is using here:


Thanks for the help everyone.


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InvisibleDoc_T
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Re: Getting started with agar, have a few questions [Re: themycoman]
    #14070040 - 03/05/11 09:12 AM (12 years, 10 months ago)

Jars are great on a small scale. But you'll want petris eventually.
Likewise with glovebox vs flowhood; I use a glovebox, but it's a bit awkward.
If I did more than a few dishes a week it would eat up a lot of time.


--------------------
You make it all possible. Doesn't it feel good?


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InvisibleJustinTime
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Re: Getting started with agar, have a few questions [Re: Doc_T]
    #14070249 - 03/05/11 10:29 AM (12 years, 10 months ago)

Slant = testube. So maybe a wire or piece of clothes hangar? Dunno.


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InvisiblePeterthinks
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Re: Getting started with agar, have a few questions [Re: JustinTime]
    #14072949 - 03/05/11 09:21 PM (12 years, 10 months ago)

Just use the tip of a syringe, you can cut agar wedges with it, spear them to move them, scrape clone tissue and they flame sterilize in a few seconds.
You probably already have them too.


--------------------
Give a man a fire and he will be warm for the rest of the night.
Set a man on fire and he will be warm for the rest of his life.
NEWB NEWBIE NEWCOMER IGNORANT? QUESTIONS?
Click HERE HERE HERE HERE For detailed instructions with pictures on how to grow mushrooms. There is a lot of info on the Shroomery and this is what you need to know.


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OfflineBlueLightRain
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Re: Getting started with agar, have a few questions [Re: themycoman]
    #14075444 - 03/06/11 12:37 PM (12 years, 10 months ago)

Quote:

themycoman said:
I was thinking I could use tweezers to get a piece out of the slant and an exacto blade (sterilized in foil of course) until I get the proper tools.




I initially purchased Stamets spear-shaped scalpel. It did okay. Then one day I was digging through my drawer and found my exacto knife. Its never necessary to replace the blade (maybe after 100 transfers) and it is much sharper and more accurate for cutting through mycelium and agar than the spear. It's cheap and it sterilizes over an alcohol lamp real well. I don't use any tin foil of sorts, I simply flame it on both sides for about 6-10 seconds, then cool it on a sterile petri dish. I've learned to keep the point of the blade up because if it's pointed downward the heat travels through the metal very quickly and can be very hot. You'll learn with practice how to work with it. I simply cut a triangle or square and then spear from the side to pick it up. Works great!

The most important item on your list should be a fire extinguisher.

I don't have any comment on what you can use for the slant. The exacto may or may not fit inside it.


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OfflineCuriousGeorge83
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Re: Getting started with agar, have a few questions [Re: BlueLightRain]
    #14075822 - 03/06/11 02:13 PM (12 years, 10 months ago)

I just ordered Stamets' "favorite scalpel' - so does a scalpel actually fit into a culture slant tube?  I've got my first culture slant coming in the mail so I am curious what to use.  I like the idea of using a syringe needle, never thought of that.
IME for agar recipes, I've never had a problem with MEA with a little nutritional yeast mixed in (which I believe makes it MYA) I do 10 grams agar, 10 grams Light Malt Extract, 1 gram nutritional yeast and 500 ml water.  This fills exactly one sleeve of plates (20 dishes).  I've cultured a fair amount of different species onto this recipe and they always take off nicely.
I recommend moving towards petri dishes though they are definitely expensive unfortunately... Also if you can get yourself a nice Erlenmeyer flask to sterilize your agar in, this makes it sooo nice for pouring.  I had always used wide mouth quart jars because I had tons of them for sterilizing rye grain, and this caused me quite a mess with pouring every time.


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Offlinepepper
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Re: Getting started with agar, have a few questions [Re: CuriousGeorge83]
    #14077491 - 03/06/11 07:48 PM (12 years, 10 months ago)

What I use is a fondue fork modified that I picked up at the dollar store. I cut the end off and then flatten the end out with a hammer. I use a file to shape the end to a tip. I have 2 different scalpels and like my home made one best.



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OfflineCuriousGeorge83
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Re: Getting started with agar, have a few questions [Re: pepper]
    #14077609 - 03/06/11 08:09 PM (12 years, 10 months ago)

:thumbup: very cool...and innovative


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InvisibleMonkeyKnifeFight
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Re: Getting started with agar, have a few questions [Re: CuriousGeorge83]
    #14078389 - 03/06/11 10:11 PM (12 years, 10 months ago)

Quote:

CuriousGeorge83 said:
I just ordered Stamets' "favorite scalpel' - so does a scalpel actually fit into a culture slant tube?  I've got my first culture slant coming in the mail so I am curious what to use.  I like the idea of using a syringe needle, never thought of that.
IME for agar recipes, I've never had a problem with MEA with a little nutritional yeast mixed in (which I believe makes it MYA) I do 10 grams agar, 10 grams Light Malt Extract, 1 gram nutritional yeast and 500 ml water.  This fills exactly one sleeve of plates (20 dishes).  I've cultured a fair amount of different species onto this recipe and they always take off nicely.
I recommend moving towards petri dishes though they are definitely expensive unfortunately... Also if you can get yourself a nice Erlenmeyer flask to sterilize your agar in, this makes it sooo nice for pouring.  I had always used wide mouth quart jars because I had tons of them for sterilizing rye grain, and this caused me quite a mess with pouring every time.




'Stamet's Favorite Scalpel' works fine for slants or whatever.  Not sure if x-acto's are better but that's the scalpel I've always used and I've been happy. 

Also this may be unpopular to say but I make all my agar from the pre-made mixes.  I have poured a ton of plates and have gone through one big container of the powder.  Honestly messing with agar is my least favorite step in the process.  It's fun to isolate cultures but when I'm just keeping an isolated culture on fresh plates it's precise work and kind of boring.  So for me it's nice to just have mixed powder ready whenever I want to pour plates.


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OfflineCuriousGeorge83
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Re: Getting started with agar, have a few questions [Re: MonkeyKnifeFight]
    #14078401 - 03/06/11 10:12 PM (12 years, 10 months ago)

good to know, thanks


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Invisiblesolarity
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Re: Getting started with agar, have a few questions [Re: CuriousGeorge83]
    #14079537 - 03/07/11 02:57 AM (12 years, 10 months ago)

Quote:

CuriousGeorge83 said:Also if you can get yourself a nice Erlenmeyer flask to sterilize your agar in, this makes it sooo nice for pouring.




Absolut-ly!
This has a good thick base that I find retains the heat and keeps the agar at that still liquid but hot enough to handle point and is then still warm (and flat) enough to sit on the stack to prevent condensation when cooling. It even has an autoclavable lid!
The Vodka's not bad either.


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