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Offlinepacmanbreed
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Re: Cleaning dirty cultures with tea agar [Re: Bigbadwooof] * 1
    #24547646 - 08/12/17 04:22 AM (6 years, 5 months ago)

thanks for sharing the recipe for tea agar.
im gonna try this soon isolating from bacteria before doing slants.
since i cant source gentamicin from our area


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OfflineFerather
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Re: Cleaning dirty cultures with tea agar [Re: Bigbadwooof]
    #24547669 - 08/12/17 05:03 AM (6 years, 5 months ago)

Quote:

Bigbadwooof said:
But what concentration of tea? Normal drinking strength?

I believe I read one poster steeped 25g in a quart worth of water. It seems quite excessive to me. Then I read your post, which was I think 6g to 250ml (about the same)? What is the effective range. I assume it's preferable to stay on the lower end of that range, and I'd rather drink my tea :wink:




I posted two recipe's for the T-Gel agar, one at 100% one at 50%, I am also using very cheap tea (60p for 250g).

"Extract: 3g (1 bag) Black tea + 125g CaCO3 water, check the end pH." -- So 1 bag to 125g water.


@pacmanbreed, Good luck m8.


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OfflineBigbadwooof
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Re: Cleaning dirty cultures with tea agar [Re: Ferather]
    #24547830 - 08/12/17 08:31 AM (6 years, 5 months ago)

Quote:

Ferather said:
Quote:

Bigbadwooof said:
But what concentration of tea? Normal drinking strength?

I believe I read one poster steeped 25g in a quart worth of water. It seems quite excessive to me. Then I read your post, which was I think 6g to 250ml (about the same)? What is the effective range. I assume it's preferable to stay on the lower end of that range, and I'd rather drink my tea :wink:




I posted two recipe's for the T-Gel agar, one at 100% one at 50%, I am also using very cheap tea (60p for 250g).

"Extract: 3g (1 bag) Black tea + 125g CaCO3 water, check the end pH." -- So 1 bag to 125g water.


@pacmanbreed, Good luck m8.




CaCO3 = Soda water?

What if I just used tap water? I never checked the end pH, just hoping for the best. It solidified alright, as far as I can tell.

Sorry to bother you with so many questions. I ran out of gent, and this seems to be a preferable alternative anyway. Just want to make sure I'm understanding everything correctly, before I start wasting plates.

Thanks for the ratio of tea:water, btw.


--------------------
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OfflineFerather
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Re: Cleaning dirty cultures with tea agar [Re: Bigbadwooof]
    #24548634 - 08/12/17 04:19 PM (6 years, 5 months ago)

No "Ca" means calcium, not sodium. Too much sodium will kill or inhibit mycelium.
Other names: Chalk, Limestone, Oyster shell, Egg shell, Agricultural lime.

Ideally you want powdered forms, you can use a shaker on grit.



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OfflineBigbadwooof
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Re: Cleaning dirty cultures with tea agar [Re: Ferather] * 1
    #24548764 - 08/12/17 05:31 PM (6 years, 5 months ago)

Oh, I have a bag of agricultural lime, and oyster shells. Sounds good!


--------------------
"It is no measure of good health to be well adjusted to a profoundly sick society," - Jiddu Krishnamurti
FARTS
"There is no need for conspiracy where interests converge" - George Carlin
Every one of you should see this video.
"If you bombard the earth with photons for a while, it can emit a roadster" - Andrej Kerpathy


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Offlinepacmanbreed
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Re: Cleaning dirty cultures with tea agar [Re: Bigbadwooof]
    #24580409 - 08/26/17 05:14 AM (6 years, 5 months ago)

Gathered my ingredients today and want to try this for cleanin my 1st cube culture asap from bac.
Thank you for sharin ur effort and deep research brother ferather.

1.)[Q]Can I use pcied dogfood agar with added tea for cube cleanin?
We cant source LME from our area.
I do understand that pure tea will work on wood lovers that produce laccase for lingin.
Im still diggin on it. Tryin to Absorb good info from good fellow to also pass it someday.

2.)[Q]being dung lovers, can cube mycelium climb up in toothpicks to lessen my vectors from scalpel?
And is it advisable vs wedge on cubes when isolating sectors?

Hope you dont mind my noobish [Q].



Edited by pacmanbreed (08/26/17 05:33 AM)


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OfflineFerather
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Re: Cleaning dirty cultures with tea agar [Re: pacmanbreed]
    #24580467 - 08/26/17 06:21 AM (6 years, 5 months ago)

1) I cant see why not, others are using standard recipe's for cubes and adding tea, with success.
2) Cubensis mycelium will happily populate a wood toothpick, for transfer or isolation.

The wood toothpicks will absorb, uptake soluble nutrients from the agar.


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Edited by Ferather (08/26/17 06:44 AM)


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Offlinepacmanbreed
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Re: Cleaning dirty cultures with tea agar [Re: Ferather]
    #24580573 - 08/26/17 08:00 AM (6 years, 5 months ago)

Love ur deep researched agar recipes brother.
Currently pcing them asm.
Will gladly report with pics in few days.


Been readin post lately. And found out this few minutes ago:
Quote:

Ferather said:
The peg will also receive nutrients from all through the agar, depending on the height of the peg.
This is due to evaporation, capillary action and mycelial uptake (distribution).


still learning more of forum  function. My bad.
Glad u have patience answering repetetive questions to help the new learners.:thanx:


Also found ur c-gel recipe.
Quote:

Ferather said:
The C-Gel agar is working, faster than T-Gel agar, and about the same speed as malt extract agar.
Both the MG solubles as nutrients and CaCO3 as carbon, are working as I intended.

Growth is slightly thicker than T-Gel, however still radial as normal.

   


Amazed how it produced nice rizho strands. :ohwow:
love to try it after sourcing MG alternative and 240bgel.
Love to see this on your sig brother someday.
Will try this on  dung lovers if time permits it.

- Can refined white table sugar be the source of sucrose?


Edited by pacmanbreed (08/26/17 01:35 PM)


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Offlinedhype773
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Re: Cleaning dirty cultures with tea agar [Re: pacmanbreed]
    #24580596 - 08/26/17 08:16 AM (6 years, 5 months ago)

This is great! With fall right around the corner, I can clone some of the local oyster without having to do 1000 transfers. It doesn't have to be pc'd, but I should be able to throw them in there with the rest of my no pours, and just keep them on hand for when they are needed right?


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Offlinepacmanbreed
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Re: Cleaning dirty cultures with tea agar [Re: Mycolorado]
    #24580713 - 08/26/17 09:19 AM (6 years, 5 months ago)

Quote:

Mycolorado said:

200ml H2O
4.25g black tea (2 bags)
2g ME
4g Agar




Thanks for the recipe.
Gonna make 2 batches One with less nutrients(ME/DF).
And 50/50 tea from eatyou.


Edited by pacmanbreed (08/26/17 01:13 PM)


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OfflineBigbadwooof
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Re: Cleaning dirty cultures with tea agar [Re: pacmanbreed]
    #24581338 - 08/26/17 01:54 PM (6 years, 5 months ago)

Seems to be working quite well on a bacterial culture I've had sitting on a slant for a long time. I have finally gotten it to fuzz up, instead of oozing like drooly ice cream. It's been on that slant for a couple years too, so it's about damn time I revive it! Gentamycin wasn't even working, btw.

In fact, the fact that this prevents spore germination makes it better than gentamycin in my book. As long as I use an intermediary MEA plate before going to grain, which I always do anyway, it will help keep things clean!


--------------------
"It is no measure of good health to be well adjusted to a profoundly sick society," - Jiddu Krishnamurti
FARTS
"There is no need for conspiracy where interests converge" - George Carlin
Every one of you should see this video.
"If you bombard the earth with photons for a while, it can emit a roadster" - Andrej Kerpathy


Edited by Bigbadwooof (08/26/17 01:55 PM)


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OfflineFerather
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Re: Cleaning dirty cultures with tea agar [Re: Bigbadwooof]
    #24581616 - 08/26/17 04:42 PM (6 years, 5 months ago)

Nice feedback.

:thumbup:


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Offlinepacmanbreed
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Re: Cleaning dirty cultures with tea agar [Re: Ferather]
    #24583234 - 08/27/17 12:50 PM (6 years, 5 months ago)

Made my 2 batches 2x7.
Both are 50/50 with varying dogfood amount.

- Hope this works as 240B gelatin substitue.
Having doubt on its sugars, carbs and sodium content.
I should have excluded it 1st, since DF is protien.

Will also try it on c-gel to verify them on second transfer without DF.


(A): 50g Water, 50g Extract, 2.5g Agar, 0.2g Gelatin, 0.75g Dogfood.
(B): 50g Water, 50g Extract, 2.5g Agar, 0.2g Gelatin, 0.50g Dogfood.


Edited by pacmanbreed (08/27/17 01:00 PM)


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OfflineFerather
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Re: Cleaning dirty cultures with tea agar [Re: pacmanbreed]
    #24583431 - 08/27/17 02:29 PM (6 years, 5 months ago)

:takingnotes:


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OfflineBigbadwooof
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Re: Cleaning dirty cultures with tea agar [Re: Ferather]
    #24583678 - 08/27/17 05:19 PM (6 years, 5 months ago)

Quote:

Ferather said:
Nice feedback.

:thumbup:




Thanks :smile:

Thanks for doing the leg work on this one too! It has probably saved me a bit of headache, and $$$ (for antibiotics).


--------------------
"It is no measure of good health to be well adjusted to a profoundly sick society," - Jiddu Krishnamurti
FARTS
"There is no need for conspiracy where interests converge" - George Carlin
Every one of you should see this video.
"If you bombard the earth with photons for a while, it can emit a roadster" - Andrej Kerpathy


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Offlinepacmanbreed
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Re: Cleaning dirty cultures with tea agar [Re: Ferather]
    #24595241 - 09/01/17 06:46 AM (6 years, 4 months ago)

Quote:

Ferather said:
Has anyone tried my sucrose (carbon source) + soluble nutrients recipe, vs, malt extract agar?
It works really well for oysters, and produced strong growth, it's mildly antibacterial.


100g > CaCO3 Water, pH 7.5 - 8.2.

0.1g > Miracle-Gro Nutrients.
0.2g > 240B Gelatin.
2g > Sucrose.

2g > Agar




Just finished Cooking my shell grit water(PH 7.5)
It Will be used for c-gel.

Planing to use cgel for second transfer of my knockup 14 tgel if they successfuly colonize.
Have used:
12 tgel for cubes.
2 tgel fo wild ganoderma clone.
(Gonna post a separate thread on it)



- Will this work for MG substitute for cgel brother?


Also Dont have sucrose from sporeworks as of the moment, but have dextrose.
Im thinkering of activating yeast with table sugar for (maltose) production based on limited reading.
- Which sugar does mycelium prefer?
- sucrose, D-glucose or maltose?
(Maybe for glycogen production)

Hope you dont mind brother ferather.
apprecite doin your hard work on this,
testing various concentration with fellow shroom brothers to give a leap off for the new comers..
Really sparked my curiosity and love of growing.. hope can pass some knowledge to the little ones someday.

I still Dont have wide knowledge on nutrient/energy utilization yet.


Edited by pacmanbreed (09/01/17 08:18 AM)


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OfflineFerather
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Re: Cleaning dirty cultures with tea agar [Re: pacmanbreed]
    #24596515 - 09/01/17 04:45 PM (6 years, 4 months ago)

That's an excellent substitute for MG. In terms of which sugar does best, I have personally only tried a few.
Sucrose (table sugar, white sugar), is easy and cheap to get, dextrose monohydrate I found best.

Dextrose monohydrate, or brewers sugar, is a D-glucose. Found in home brewing.
You will also find yeast nutrient in the same section, adds vitamins.

   


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Re: Cleaning dirty cultures with tea agar [Re: Ferather]
    #24597733 - 09/02/17 07:47 AM (6 years, 4 months ago)

A note with the DIY agar recipe, it was never intended to be anti-bacterial, it's simply an alternative method.
It can be used for learning purposes as well, I guarantee vigorous mold in 3 days, be fully aseptic.

The good news is, it germinates spores beautifully, due to the soluble nature.
You can also use the same recipe, without agar for liquid cultures.

It is no different, other than DIY, to ME or PD and so on.
100% sugar and water will eventually stall.


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OfflineBigbadwooof
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Re: Cleaning dirty cultures with tea agar [Re: Ferather]
    #24599732 - 09/03/17 02:57 AM (6 years, 4 months ago)

I just transferred from the tea agar plate to a few MEA plates. I didn't see any bacterial contamination on the tea plate. The culture I am growing has been on a slant for years, and it was quite heavily contaminated with bacteria. I cut a piece of colonized wood from a popsicle stick I had in the master slant, and used it to inoculate the tea plate, which I have now used to inoculate the MEA plates.

It is worth noting that when I made the tea plate, I also made an MEA plate from the same slant. The MEA plate has two distinct drips of bacteria going all the way to the edge of the plate, like drooly ice cream, but the rest of the plate appears to have grown out alright, to my surprise. The tea plate had very faint, tomentose growth, while the MEA plate had rhizomorphic growth. You can tell the age of the culture, by the rhizomorphic growth (on the original MEA plate), as it does not look terribly vigorous. The rhizomes are sparsely spread.

My phone is broken, so I can't post pictures.

One thing that I have been doing to combat contamination, that I think a lot of people do improperly, is to transfer only a very tiny sliver of mycelium from the very outer edge of visible growth, on to the next plate. I often see people transferring large wedges, and while that may speed up colonization of the plate, it also increases the potential for transferring contaminates.

Anyways, I will keep you guys posted, if you are interested in my results. I intend to take the 3rd transfer plate and make an agar slurry to inoculate jars with. Maybe I am jumping the gun, but I am really anxious to get things going again. I also have an AA+ culture that is equally old as fuck, which is contaminated with mycogone. I saw the mycogone growing on the plate, and I am attempting to transfer away from it, but this shit seems to be very difficult to get rid of.

TL;DR: Just made transfers from tea plates to MEA. All looks good. Hoping that continues, and I will keep you all posted!

I'm getting that sort of giddy impatience I had when I first started growing, because I want to see my precious old cultures fruit once again!


--------------------
"It is no measure of good health to be well adjusted to a profoundly sick society," - Jiddu Krishnamurti
FARTS
"There is no need for conspiracy where interests converge" - George Carlin
Every one of you should see this video.
"If you bombard the earth with photons for a while, it can emit a roadster" - Andrej Kerpathy


Edited by Bigbadwooof (09/03/17 03:01 AM)


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OfflineFerather
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Re: Cleaning dirty cultures with tea agar [Re: Bigbadwooof]
    #24600121 - 09/03/17 09:43 AM (6 years, 4 months ago)

Awesome work, keep it coming.

:takingnotes:


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Growing mushrooms, general guide and information (Ferather's Journal), https://ibb.co/rG3rML2

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