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Simple agar prep


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#1 peacefrog

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Posted 01 March 2015 - 08:07 AM

I was just embarking on a new project and thought I would share my simple agar preparation for any out there that feel intimitated by agar.

This morning I decided to make one of my favorite and simple agar recipes, malt agar. I always make my own agar, but prepared agar for mushroom cultivation you can purchase is great. I have used both with the same results. I'm just a do it yourself kind of guy, and love preparing something from scratch (and it's cheaper).

I use 1/4 pint jelly jars instead of petris. IMO this saves money and time using these jars for the hobbyist like myself. Plus no need to make more then you may use, and no need for the added step of pouring.

Simple agar agar one can find in most health food stores and light malt extract found in a brewery store:
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For 5 jars I mix 1 tbls of agar agar to 1tbls of malt in a small bowl:
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I then fill the jars up with ~ 3 tbls water:
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Add ~ 1 teaspoon of the agar mix to each jar and stir, cover with foil and reay for the pc:
image.jpg image.jpg

I then sterilize at 15 psi for 30 minutes. After sterilizing, solidifying, and cooling they are ready for inoculation. Nothing new here, and I'm sure that others utilize similar means of agar work. Just thought I would share since I was doing it anyway.

Edited by peacefrog, 01 March 2015 - 10:16 AM.

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#2 hyphaenation

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Posted 01 March 2015 - 08:09 AM

Good stuff... thank you. I like those jars for agar.

#3 Headinthehills

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Posted 01 March 2015 - 08:21 AM

This is going to be my next experiment. I just got a clone going and want to make a master on agar. Thanks for the write up.

#4 peacefrog

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Posted 01 March 2015 - 08:24 AM

Thanks, Hyph.

Yea me too. Aftera few years and a lot of money spent on pre-sterilized sleeves of petri dishes, and making too much agar in a glass bottle, I decided to go in a different direction. I am no where near a commercial grower, so that just didn't make since for me to keep spending the revenue and time on that stuff for cultivating a few crops per year. Those little jars don't give you the same amount of surface area as a petri, but they are more than adequate for agar work IMO.

#5 PsyBearknot

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Posted 01 March 2015 - 09:13 AM

Very nice write up and VERY simple process!

My I suggest you edit your original post and add the PC time and PSI. I Think that would make it more of a complete picture to some one just starting out.

Nice job man!
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#6 happy4nic8r

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Posted 01 March 2015 - 06:22 PM

I am trying something very similar, and have a couple of questions, if you don't mind.

 

I like the way you don't cook the agar, the pc just mixes and makes it uniform, with no stirring?

 

Are the lids  put on upside down, or are they banded two piece canning lids?

 

I still have a half dozen that I did with the lids like regular canning, vacuum sealed as it were by the pc, and 4 days later they still have nothing growing on them.

 

The ones i opened up for any reason either have what I put on them growing, or white film all over surface, or both. The jars worked much better than the plates I made with them.

 

never mind what I did,

 

I want to do it right, so I'm glad you posted this, very simple recipe.

 

I had extra water somehow in the mix as well and it condensed out and just sits there.

 

Thank you Peacefrog, nice and concise!!



#7 peacefrog

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Posted 02 March 2015 - 03:41 PM

Happy,

Yes I stir in the water once the agar is added. The pc does the rest of the work. And yes all bands are upside down just like you would do with cakes.

And some condensation is to to be expected but isn't really an issue for me. There is a trick to keeping the condensation from dripping onto the surface. When inoculating or streaking spores, very carefully open the lid and tilt it to the side away from the agar surface. Most if not all will drip harmlessly away onto the surface of where ever you are working. Some water will inevitably drip. Usually will not be an issue. Good sterile work and proper sterilization and you should be fine. As always, our enemy will find a way to take over sometimes. Just happens.

#8 Chu

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Posted 02 March 2015 - 07:58 PM

Great stuff... I've used half-pint sized jars before, then I bought a 400 petri-dish case (what a fool) which has been moving with me for the past 12 years... one day they'll end and I'll go back to jars and slabs only...

 

What's up with that "lids upside down" thing... I've ever used it the right way, with the rubber ring down...

 

Thanks



#9 peacefrog

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Posted 02 March 2015 - 09:34 PM

The rubber side down will work too. But if you leave it down the lid tends to seal, making it harder to remove after sterilizing. Leaving the rubber seal up allows easier access for inoculations.
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#10 PsyBearknot

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Posted 03 March 2015 - 04:13 AM

I think not only harder to remove but when the seal is broken there is a difference in pressure that draws air into the jar. Not an issue if your in a clean space but still an added risk that is un necessary.

Edited by PsyBearknot, 03 March 2015 - 04:13 AM.

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#11 Chu

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Posted 03 March 2015 - 07:52 AM

Shit... everything is obvious... after someone tells you about it... of course! My lids did stick... but I wouldn't think about using them in any other way than the directions on the box...

 

Great tip, many, many thanks!



#12 proZac

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Posted 04 March 2015 - 01:42 PM

Very nice post :-). I like to make my own stuff too:-). What do you think guys? Can I make or theres any point to do a following tryout?

-boil straw / rye to an hour
-filter the solution
-fill any type of jar you like
-pc-ing
-add agar,dextrose ...

Question is...

Can it replace the malt extract with a good efficiency?

IMO if I will use grain spawn after the cloning session the myc more accessible with grain :-). Is it possible?

#13 happy4nic8r

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Posted 04 March 2015 - 03:03 PM

You can make agar a thousand different ways, and at least half of them will work, some work better than others.

 

It has been my limited experience that this has a "weakest link" type embedded glitch.

 

One wrong move, or a sneeze can introduce fast growing contams, unlike anything I've seen on other substrates.

 

I bought pre filled plates, some sterilized plastic, and some sterilized glass.

 

The pre-filled quickly grew some kind of bread yeast mold.

 

The glass I tried to make and sterilize a different kind of agar, it got a white slimy bacteria on it, before either the multi spores, or the agar transfer had a chance.

 

I cleaned it all up and tried again, this time with jars added to the mix.

 

The plates again got the white bac, the jars worked perfect, so far. It has been less than a week, and the agar transfers (all I did this time with azures), are all spreading slowly into the agar.

 

The jars I left sealed, (lids rubber side down), are still good to go, nothing growing in them, and there shouldn't be if my SAB is working correctly (and no sneezing).

 

I don't recall any agar recipes calling for straw, but I think to add some nutrients like yeast or potato would be easier.

 

I like peacefrog's recipe here, just simple sugars and agar. They are both pretty cheap and easy to get. I added a touch of yeast, but you don't have to.



#14 Chu

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Posted 04 March 2015 - 07:54 PM

Hey, I use, per liter:

 

20g Malt Extract

10g Dextrose

5g Yeast Extract

3g Meat or Casein Peptone

 

After a few years I found that adding the Meat or Casein Peptone speeds up growth considerably for hyphae.  

 

Other than that, I've tried several other recipies which involved making a broth out of something... Popcorn, WBS, Rye, Wheat, red meat, egg white, milk, etc... none of them came even close to using the standard recipe I mention above. I've tried making the Potato agar (PDA), but still found the above a lot better.

 

Fungus will grow on anything, even on water alone (for a short time), so you can use pretty much any food source to grow it. The question is, how can one optimize the process. In this case, it seems like speed of growth and duration of nutrition would be the keyparameters... but if access to the ingredients is an issue to you, then you can always compromise on speed and duration in order to work with what you have.

 

Anything will work if made and kept sterile.

 

Have a good night fellas!



#15 Chu

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Posted 04 March 2015 - 07:55 PM

Sorry... I've of course failed to mention something on the receipe above... it's for Liquid Culture... same recipe for agar, I just add 10g per Liter of the one I currently use.






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