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Agar experimenting


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#41 Microbe

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Posted 25 November 2014 - 12:48 PM

Nice!!! What mushroom is this mycelium from?

Dont know. I call it Tex lol. It was supposed to be PE bit afyer fruiting it it clearly was not and i had a few say it could be PE6, texans, and maybe 1 other but i dont remember. Someone also told me it xould have reverted back or reversed the mutation so would that make it PE6? Im just going to call it Tex for now. :)
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#42 Microbe

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Posted 27 November 2014 - 10:18 AM

Plate 1-1 went in the fridge lastnight as it srarted to metabolize the agar. I will end up isolating it on agar before fruiting. I only need a few more transfers IMO before i obtain a isolate or monoculture.

The remainder of the plates are going into cold storage as well today until i get around to working with them. I will conduct another transfer on each of the 3 plates and then fruit and isolate from there via clone but i more then likely will not continue forward with 1-2, 1-3, and 1-4 after fruiting unless i see amazing fruiting results. I dont see the need the need to have more then just a few isolates laying around. Anyway here are the pics and i would consider this experiment a success.

7378f6b57471258471a66ec48ced7acd.jpg

98a401678a15053befb6cca56bf2f400.jpg

6e0ac76725da3428b885c5d354aad8aa.jpg

Not completely rhizo growth across the plates but I would put any 3 of these cultures into grain jars ;)

Edited by Microbe77, 27 November 2014 - 10:19 AM.

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#43 TurkeyRanch

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Posted 27 November 2014 - 10:53 AM

Great thread.

archive material.

Digging the black agar! Looks like your having success.
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#44 whitethumb

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Posted 28 November 2014 - 04:41 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/
How many half pint jars does 125 ml of water and 5 gm of mea make?

#45 Microbe

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Posted 28 November 2014 - 11:38 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/
How many half pint jars does 125 ml of water and 5 gm of mea make?
Depends on how thick you pour your agar. I would estimate 10 assuming in takes about 15 ml to give you a thin layer. If you plan on using them soon then pour as thin as you can so you can get more mileage out of your agar.

#46 happy4nic8r

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Posted 06 December 2014 - 02:28 PM

I am loving this thread, and the other one that referenced this one. I'm checking on supplies today, I can give them to my wife as Xmas presents, and she'll go, what the fuck??? This should definitely be archived, great pictures, great recipe. Way to go Microbe77!! Would love a follow up on the continuing saga...some of your other technique. 


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#47 whitethumb

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Posted 13 December 2014 - 11:11 AM

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/
Just any kind of ziplock bags?

#48 Grey_Being

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Posted 13 December 2014 - 05:01 PM

Nice Post Microbe  :wub:

Agar really takes the world of mycology to a whole new level, and separates the hobbyists from the dedicated! The one true way to 

achieve the end results one truly desires consistently! 

BTW, where did you get that nifty flow hood your working with? Ive been thinking of a way to design and have an affordable model made

by outsourcing to someone with a 3D Printer, although it would be much cheaper than say a 3000$ lab model, it still seems it may add up 

quick! So Im just curious where I can get one of those, Much appreciated  :biggrin:


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#49 torn2bits

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Posted 13 December 2014 - 10:07 PM

Great work, this IMHO should stay separate from your other agar thread.
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#50 Needles

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Posted 14 December 2014 - 01:13 PM

Great effort to control your mycelium there microbe. Agar work is my favorite part of mycology. Isolating sectors to get the strongest strain is the way to go. This has been a good thread, hopefully it encourages others to go to work on their own agar projects. Once you get the hang of it and have your sectors isolated or even a clone or two . You will have so many more opportunities to excel in growing mushrooms. more than you would with just a spore syringe and pf cake. Adding the black food coloring is a very cool twist, can't wait to give it a try.
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#51 Microbe

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Posted 14 December 2014 - 03:13 PM

Nice Post Microbe :wub:
Agar really takes the world of mycology to a whole new level, and separates the hobbyists from the dedicated! The one true way to
achieve the end results one truly desires consistently!
BTW, where did you get that nifty flow hood your working with? Ive been thinking of a way to design and have an affordable model made
by outsourcing to someone with a 3D Printer, although it would be much cheaper than say a 3000$ lab model, it still seems it may add up
quick! So Im just curious where I can get one of those, Much appreciated

I built it and it maybe cost 600 including the cabinet it is mounted to. I didnt document it but i just framed in around a 24×24×12 filter, the kind used in air scrubbers, not the most efficient but it works and cost just a little over 200.00.

Framed around the filter with MDF leaving a 8" plenum and a 10" working surface away from the filter. The blower is a centrifugal blower wired to a rheostat. I bought in on clearance at amazon and is over powered so i will never have to worry about loosing laminar flow as the filter becomes clogged, at least not in the next few decades.

Some must do's IMO

I cleaned the inside of the box with bleach solution and allowed it to be absorbed into the MDF then applied a very generous amount of shellac finish. This should prevent any mold from growing and colonizing inside the plenum.

I stack 2 filters, the first being a allergen filter i bought on clearance, and then the top is a polyfill netting type filter.

A flowhood is not required for agar work and a Still Air Box can provide a equally effective sterile enviroment when used properly. Flowhoods are just more ergonomical.
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#52 Microbe

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Posted 14 December 2014 - 03:21 PM

Great effort to control your mycelium there microbe. Agar work is my favorite part of mycology. Isolating sectors to get the strongest strain is the way to go. This has been a good thread, hopefully it encourages others to go to work on their own agar projects. Once you get the hang of it and have your sectors isolated or even a clone or two . You will have so many more opportunities to excel in growing mushrooms. more than you would with just a spore syringe and pf cake. Adding the black food coloring is a very cool twist, can't wait to give it a try.

I too feel that agar is my favorite step in mycology. Its not difficult and there are many layers and reasons to why agar work is conducted but IMO basic agar work is no more advanced then a PF Tek. If you look at all the steps from start to finish on a PF Tek, or any other tek at that matter, agar is not difficult. You have to develop a sterile procedure and then the rest is developing a trained eye. If you can successfully colonize a jar of grain, you can do agar work.
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#53 Microbe

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Posted 14 December 2014 - 03:24 PM

Great effort to control your mycelium there microbe. Agar work is my favorite part of mycology. Isolating sectors to get the strongest strain is the way to go. This has been a good thread, hopefully it encourages others to go to work on their own agar projects. Once you get the hang of it and have your sectors isolated or even a clone or two . You will have so many more opportunities to excel in growing mushrooms. more than you would with just a spore syringe and pf cake. Adding the black food coloring is a very cool twist, can't wait to give it a try.

I too feel that agar is my favorite step in mycology. Its not difficult and there are many layers and reasons to why agar work is conducted but IMO basic agar work is no more advanced then a PF Tek. If you look at all the steps from start to finish on a PF Tek, or any other tek at that matter, agar is not difficult. You have to develop a sterile procedure and then the rest is developing a trained eye. If you can successfully colonize a jar of grain, you can do agar work.
And adding black food coloring to agar makes a HUGE diffrence and i think any who try it will never use regular colored agar.
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#54 mydarling

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Posted 14 December 2014 - 07:38 PM

Cool idea using black food coloring. I've never seen that one before ... just various colors due to nutritive/antimicrobial additives. But black is cool! Makes the hyphae pop.

 

I also like the information you've got here about SABs. Flowhoods are nice but you can certainly work without one. Thanks for sharing.

 

Nice contributions here. Yay agar!


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#55 Grey_Being

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Posted 20 December 2014 - 03:47 AM

Cool Thanks for sharing Microbe I've always dreamed of having/building one. I guess all the years I had access to one one, I never thought to relish in it and ever since I have thought of ideas to construct one to pursue the work our mother earth has laid upon us to share the knowledge and keep her creations alive!!!   :meditate:



#56 peacefrog

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Posted 20 December 2014 - 07:57 AM


Great effort to control your mycelium there microbe. Agar work is my favorite part of mycology. Isolating sectors to get the strongest strain is the way to go. This has been a good thread, hopefully it encourages others to go to work on their own agar projects. Once you get the hang of it and have your sectors isolated or even a clone or two . You will have so many more opportunities to excel in growing mushrooms. more than you would with just a spore syringe and pf cake. Adding the black food coloring is a very cool twist, can't wait to give it a try.

I too feel that agar is my favorite step in mycology. Its not difficult and there are many layers and reasons to why agar work is conducted but IMO basic agar work is no more advanced then a PF Tek. If you look at all the steps from start to finish on a PF Tek, or any other tek at that matter, agar is not difficult. You have to develop a sterile procedure and then the rest is developing a trained eye. If you can successfully colonize a jar of grain, you can do agar work.
Amen. Imo, agar is the way to go even from the start. It is very easy and will open up a lot of doors for your cultivation endeavors. And I also like the addition of black food coloring. Never thought of that, but I often use h-poo water in my agar plates for dung lovers and the dark color does help you. Nice thread, Microbe.

Edited by peacefrog, 20 December 2014 - 07:59 AM.





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