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OfflineFerather
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Testing nitrogen sources with agar [Quick guide]
    #24962441 - 02/02/18 07:28 PM (3 years, 3 months ago)

How to test nitrogen sources using plain sucrose as the carbon source (no nitrogen -- no protein, no vitamins).

100g - Boiling hot water > 2g - Agar powder > 2g - Sucrose > 0.024g - Nitrogen.

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In my case I would add 0.1g MG soluble nutrients (24%-8%-16%).
The setup will grow pretty much anything so be aseptic.

   

The MG also dyes the agar blue.


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Invisiblenube424
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Re: Testing nitrogen sources with agar [Quick guide] [Re: Ferather]
    #24963369 - 02/03/18 04:33 AM (3 years, 3 months ago)

Ive been wondering this. Does mg help growth in any way?


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OfflineFerather
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Re: Testing nitrogen sources with agar [Quick guide] [Re: nube424]
    #24963636 - 02/03/18 10:08 AM (3 years, 3 months ago)

It depends on the mycelium, if the mycelium can use ammoniacal and-or ureic nitrogen as the sole source, yes.
Oyster's and Cubensis will utilize both as the sole source of nitrogen, I cannot say for other's.

MG also adds other "essential" macro-micro nutrients, which are also utilized.
Ignore the plant label, just see the soluble nutrients, and usability.

----

MG or similar, should only be used on plain substrates.

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Here is the readily available Oyster data (I have also tested it on Cubensis).

Carbon sources (sugars): https://www.ncbi.nlm...97375/table/T3/
Nitrogen sources: https://www.ncbi.nlm...97375/table/T4/

Original paper: https://www.ncbi.nlm...les/PMC4397375/


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Edited by Ferather (02/03/18 11:22 AM)


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OfflineFerather
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Re: Testing nitrogen sources with agar [Quick guide] [Re: Ferather]
    #24998134 - 02/16/18 08:24 AM (3 years, 2 months ago)

First test:

Recipe: 50g water, 1g agar, 0.5g sucrose, 0.02g MG. End pH: 8.


At pH 8 sucrase enzymes are about 40% efficient. 0.5 - 60% = 0.2g sugar (0.4%).

My sample is carbon weak, however it's not stalling, and structured.
I need to acidify the agar media, without adding nutrients.

   

Any cheap suggestions for an acid?


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Invisiblenube424
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Re: Testing nitrogen sources with agar [Quick guide] [Re: Ferather] * 1
    #24998256 - 02/16/18 10:30 AM (3 years, 2 months ago)

Vinegar?


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OfflineBuster_Brown
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Re: Testing nitrogen sources with agar [Quick guide] [Re: nube424] * 1
    #24998277 - 02/16/18 10:38 AM (3 years, 2 months ago)

Acid rain?


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OfflineFerather
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Re: Testing nitrogen sources with agar [Quick guide] [Re: Buster_Brown]
    #24999247 - 02/16/18 06:50 PM (3 years, 2 months ago)

Not sure about vinegar, it contains acetic acid, which can kill things at 3%.
If it works for pH, that would be interesting and very beneficial.

I will try with vinegar to get pH 6.0-6.5 or around.


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Re: Testing nitrogen sources with agar [Quick guide] [Re: Ferather] * 1
    #24999261 - 02/16/18 06:57 PM (3 years, 2 months ago)

Quote:

Ferather said:

Any cheap suggestions for an acid?




https://en.wikipedia.org/wiki/Hydrochloric_acid
Quote:

pH control and neutralization

Hydrochloric acid can be used to regulate the acidity (pH) of solutions.

        O H −  +  H C l  ⟶  H 2 O  +  C l − {\displaystyle \mathrm {OH^{-}\ +\ HCl\ \longrightarrow \ H_{2}O\ +\ Cl^{-}} } {\displaystyle \mathrm {OH^{-}\ +\ HCl\ \longrightarrow \ H_{2}O\ +\ Cl^{-}} }


In industry demanding purity (food, pharmaceutical, drinking water), high-quality hydrochloric acid is used to control the pH of process water streams. In less-demanding industry, technical quality hydrochloric acid suffices for neutralizing waste streams and swimming pool pH control.[26]




you can get it at any hardware store or pool supply as muriatic acid.


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OfflineFerather
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Re: Testing nitrogen sources with agar [Quick guide] [Re: ballsalsa]
    #24999291 - 02/16/18 07:15 PM (3 years, 2 months ago)

Thanks mate.

:takingnotes:


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Re: Testing nitrogen sources with agar [Quick guide] [Re: Ferather]
    #24999433 - 02/16/18 08:02 PM (3 years, 2 months ago)

On second thoughts, I actually have two options:

Option 1: Add more sucrose, and overcome deficiency via content.
Option 2: Add HCl to increase efficiency of enzymes.

One is cheaper than the other (option 1), however I cannot decide which would be worse.
Adding more sucrose and increasing efficiency both add to contamination level.

What do you think is the better option, and why do you think so?


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Re: Testing nitrogen sources with agar [Quick guide] [Re: Ferather]
    #24999479 - 02/16/18 08:19 PM (3 years, 2 months ago)

depends on if you think the mycelium would be ok at even higher ph.  If so, then maybe you could eliminate some contams that way and be safer by adding more sugar.  Or you could play with other organic carbon sources like ethanol.


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Re: Testing nitrogen sources with agar [Quick guide] [Re: ballsalsa]
    #24999509 - 02/16/18 08:34 PM (3 years, 2 months ago)

I was thinking about replacing sucrose as you have mentioned, previously I have used CaCO3 (converts to bicarbonate).
The bicarbonate and MG produced a full plate, however CaCO3 stops dissolving ~pH 9, not enough is made.

The CaCO3 based carbon source ended up similar to the growth I have with sucrose, quite weak.

----
----

Results:

I conclude the added nutrients are being utilized as intended with my carbon source.
Mycelium require nitrogen and other macro-micro nutrients with basic carbon.

----

Next steps:

Find another viable carbon source, add more sucrose or change the pH.


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Re: Testing nitrogen sources with agar [Quick guide] [Re: Ferather]
    #25000387 - 02/17/18 04:38 AM (3 years, 2 months ago)

Carbon source... Charcoal?


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Re: Testing nitrogen sources with agar [Quick guide] [Re: nube424]
    #25000531 - 02/17/18 08:00 AM (3 years, 2 months ago)

Mycelium wont cleave the C=C bonds, the trace ash it will though, but that's the same as CaCO3.


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Re: Testing nitrogen sources with agar [Quick guide] [Re: Ferather]
    #25001674 - 02/17/18 06:40 PM (3 years, 2 months ago)

I will do a higher sucrose content when I next test it again. Here is T-Gel plain, and T-Gel + MG.

               

The population rate and potential with MG was much higher than without.


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