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Agar experimenting


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#21 TurkeyRanch

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Posted 01 November 2014 - 08:41 PM

I use the site specific search on google,

site:mycotopia.net how to pasteurize 50 lbs of anything
site:mycotopia.net flowhood
site:mycotopia.net glovebox

This way you only get results from Mycotopia.
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#22 torn2bits

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Posted 03 November 2014 - 06:07 AM

Micro....
Tell us about these lights in the still air box...
I see power cords so what type of lights are they man?

#23 Alpine

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Posted 03 November 2014 - 04:00 PM

You two are inspiring me to break my agar "seal".

When I made my first few attempts to make and pour agar it was a complete failure.... Some plates wouldn't harden due to a bad mix or some would contaminate. BUT THEN I bought this stuff on eBay for $15 (mea agar premixed) and it kick started this noob into the world of agar! There is 114g of mix in it and instructions say 20g makes 500ml of agar so.... I take 5g and microwave some Distilled water to almost boiling then carefully measure 125ml of it into another jar then add the 5g of mea then screw a lid on and vigorously shake. (The malt mixes right in at this point but the agar won't completely mix until PCed so it will look a little cloudy FYI) Give the jar a good shake and use a 60ml syringe to transfer 10-20 ml to each wide mouth half pint jar or PCable petri dishes or any other heat resistant container desired. Then PC as instructed on label (When Iv used jars I would poke a hole in the lid and cover with tape for later inoculation then place the jar in a ziplock n cover with foil then PC. The ziplock will keep moisture out during PC but is also a fancy way to store the agar jars until needed for innoc)

This Tek posted by Hyphaenation adds to this quite well. Check this out.
There are also more useful links posted within this link check those out to!

Redboy -potato/malt agar jars - G2G / Myc Syringes - Mycelium Milking / Super-Inoc
https://mycotopia.ne...ing-super-inoc/

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Edited by Alpine, 03 November 2014 - 05:32 PM.

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#24 CatsAndBats

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Posted 03 November 2014 - 05:00 PM

very thorough. thank you



#25 Alpine

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Posted 03 November 2014 - 06:14 PM

No problem!. I like hyphs idea of adding the popcorn to the jars for Myc milking or g2g.

Also if there is anyone that hasn't experimented or prepped popcorn yet here is an awesome tek posted by Irishlion.

Popcorn Simple and easy with single hole polyfil lids
https://mycotopia.ne...ds/#entry831402

But for the first step instead of prepping a whole pressure cooker full of popcorn.
Add a 1/2 cup of popcorn per wide mouth pint jar then add 240-250ml water (basically fill up to the line below the threads of the jar) pop 4 large holes in lids and tighten metal side down and wrap tight in foil then PC 15 psi for 60 min. Then follow the rest of the tek I linked for final steps and sterilization.

Edited by Alpine, 04 November 2014 - 12:34 AM.

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#26 Alpine

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Posted 03 November 2014 - 06:41 PM

.Sorry for posting a little off topic
I actually plan on posting that info and links in a good place for newbies to find.

I like your glove box Microbe It's inspiring me to think more out side the box on my own setup. Maybe you could post a few pix of it and how your arm holes are setup and what kind of lights your using. (The light I tried on mine made it to bright outside the box n made it hard to see the flame for the needle) Keep us updated on your experiment!
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#27 Heirloom

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Posted 12 November 2014 - 06:00 PM

I have decided that I can do agar, am waiting on mea. using 4 oz mason jars the
smallest made, about 50 mm @ the bottom.

 only doing a few exotics , ps. cyans to nocc up sub.

    I believe that after all the reading I have done I can do it with my still air box.

      I advance a little at a time, growing in confident knowledge, thanks to the teachers here.


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#28 Microbe

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Posted 13 November 2014 - 01:40 AM

Micro....
Tell us about these lights in the still air box...
I see power cords so what type of lights are they man?

Puck lights.

#29 Microbe

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Posted 13 November 2014 - 01:53 AM

.Sorry for posting a little off topic
I actually plan on posting that info and links in a good place for newbies to find.

I like your glove box Microbe It's inspiring me to think more out side the box on my own setup. Maybe you could post a few pix of it and how your arm holes are setup and what kind of lights your using. (The light I tried on mine made it to bright outside the box n made it hard to see the flame for the needle) Keep us updated on your experiment!

I will get some detail pics. Its prettybasic though. I purchased 2 6" pvc couplers traced the circumference and cut the holes out then siliconed them in. The pic didnt show it as that was an old one that i showed but i also use the test caps that a screwed cup hooks so that i can plug the arm holes up when not using it. I also cable tied a few pieces of tyvek to the top of the pvc couplers so that when i remove my arm or arms from the SAB the tyvek flaps fall down over the opening. The lights are just under cabinet lighting i installed. I rarely use since i built my flowhood.
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#30 Luckyloser

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Posted 13 November 2014 - 06:28 AM

I've actually got all the stuff to make a SAB just like that (had plans to but never did). I didn't think that I would be that successful with agar but no hood. If you were just starting out with agar, would you think a SAB would better than a shmuv box or the opposite. I'm wanting to get into agar but don't want to waste my time nor do I have the space or funds for a hood. Thanks in advance.

#31 Microbe

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Posted 14 November 2014 - 11:39 AM

I've actually got all the stuff to make a SAB just like that (had plans to but never did). I didn't think that I would be that successful with agar but no hood. If you were just starting out with agar, would you think a SAB would better than a shmuv box or the opposite. I'm wanting to get into agar but don't want to waste my time nor do I have the space or funds for a hood. Thanks in advance.

I started in a SAB. The only reason i built a flow hood was for ergonomics . Its sucks working with 20+ spawn jars trying to do g2g. But agar work can be done successfully in a SAB especialy when working with contaminated cultures. When used properly a SAB will provide just as sterile working enviroment as a flow hood. Heck it might even be better as with flowhoods you still have some particles that make it through the Hepa. These particles exit the filter face in laminar flow so they shouldnt drop down onto your working surface. The smuvbox IMO is a no go for me. The problem i see with the design is that the hepa filter will allow particulates through and inside a plastic bag they are swirling around everywhere. I know many use them and with success but i have successfully conducted g2g in open air on a countertop in my kitchen.

The concept behind the SAB is that when you spray the walls with a dish detergent solution, i prefer to use bleach solution, any particles are trapped in the solution stuck to the walls and ceiling of the SAB so that they dont drop into your shit. You spray the air with a lysol, which i also use bleach solution for this, and this helps pull any particles floating around to the floor of the SAB. You wait 10 minutes for anything else to settle before you begin your work. It is best to never have to withdraw your arms once you start so dont use anything flammable in your SAB or you will have to flame steralize outside of your SAB which is ok i did it all the time but each time you withdraw and insert your arms you could be bringing in contaminated air.

You will be Absolutley fine conducting agar work in a SAB.
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#32 Heirloom

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Posted 15 November 2014 - 01:18 PM

the concept behind using a sab amounts to simple instructions and reminders to others.

 a person really needs a dedicated room, to work in , grow in ..ect plus to store " tool " .

 thanks


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#33 Alpine

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Posted 16 November 2014 - 05:21 AM

Microbe, Are you still working on your agar experiment? If so how's it going?

#34 Heirloom

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Posted 18 November 2014 - 06:17 PM

I'm glad I read this thread. I bought some MEA and now am going to use it to
try to save some azure cultures, that looks like a little contams.
also to start some p. cyans  to spawn 


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#35 Microbe

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Posted 20 November 2014 - 03:42 PM

Microbe, Are you still working on your agar experiment? If so how's it going?

Getting ready to start transferring in the next day or two. I have the plate in cold storage right now. Holidays coming up, christmas shopping for 5 kids, and etc has delayed me a bit. But yeah im going to play it out and see what happens. I will update when i do.

Edited by Microbe77, 20 November 2014 - 03:42 PM.

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#36 Microbe

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Posted 22 November 2014 - 03:28 PM

I wanted to make 4 transfers from this plate. 1 of which were taken from an area where there were no visible signs of bacteria and 3 of which i selected the bacteria with the myc.
5e93cfa0587a9d31db62330ef8984f78.jpg

Plate after being in the fridge for a few weeks.

b2db35a84d3caa53f2867dcc8aaf21e4.jpg

My first selection 1-1 away from the bacteria. This had some rhizo growth that was clearly visible before going into the fridge. I typicaly never lrt my plates grow out to the edge unless im increasing inoculant.
81889be6cbbf5578aa8e184e760f3b68.jpg

My second second selection 1-2 showed rhizo growth overtaking the bacteria.
f34fc300e3094e8bfa30734fd1f8495d.jpg

My 3rd selection 1-3 showed rhizo growth overtaking the bacteria.
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My 4th selection showed some really aggressive rhizo growth and was very easily and rapidly overtaking the bacteria.
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Its hard to see the rhizo growth as the cold storage thinned it out alot but i assure it is there and we should see it on the plates hopefully when they start colonizing. I will share the results.
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#37 Microbe

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Posted 23 November 2014 - 12:46 PM

Should this be combined with with the agar work thread i started or should it stay right where it is at?

#38 Microbe

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Posted 24 November 2014 - 09:39 AM

UPDATE

Sorry for the horrible pics. My ziploc bags, and papertowel residue are making it hard to capture a good one. Maybe i will start using

Nice rhizo growth starting. No signs of bacteria.

915c7b0bd8213de4be23dbc96ba38260.jpg

Another plate starting to show rhizo with no signs of bacteria.

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Its hard to tell from the picture but there is some bacteria around the base of the agar plug and also there is some rhizo growth starting to show. Myc.

66fd5696a500f70f6789fe8f4c0d77b7.jpg

Might be hard to tell from this plate but it also has bacteria showing and there also is some rhizo growth. This specimen was selected right from the front lines of the myc and bacteria war. Dont let the air bubbles fool you as that is all it is and it happen while aspirating the last bit of agar from my syringe and the agar solidified trapping the air pockets. This is the plate that will give me solid evidence that the substrain im working with will compete very well with this unknown bacteria, at least on agar anyway.

760367ea17401f6d571b374530eba57a.jpg

I will keep everyone updated.
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#39 Microbe

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Posted 25 November 2014 - 12:42 PM

UPDATE

Im helping wife put up tree, Christmas lights, and all the other holiday fun stuff and i have no plates poured so 1-1 is probably going in the fridge until the upcoming weekend. But so far they are all looking good.

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#40 whitethumb

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Posted 25 November 2014 - 12:45 PM

Nice!!! What mushroom is this mycelium from?




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