anders said:

ThurmanMurman said:

My brother in Christ...

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Ja Thurman, I understand your true intentions here, and as an aside although I do believe in the teachings of the leader Christ, almost no one these days understands nor actually knows them, much less really lives by them.  And in any case I am an atheist.

Your heartfelt post is enough for me to change my practice of excising trich and other contaminents, to excising the healthy instead, as I am not intransigent, and I respect you.  Science makes the decision.

Also I have made further developments in this craft, but feel unsafe in this environment so I will practice them on my own.  (A Mod, who I did not know is a Mod, is allowed to insult and punish us, however we are not allowed to respond in kind?)  Well it's the way it is. 

I keep my developments to myself.  It is my consolation.

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Headass said:

I did get some TSA, but have not poured it yet.  I just got to get it poured

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Quote:

Bunny.12 said: Hi, thank you for the information. I just bought the syringe of spores as it was one of the few places selling material in my country. I had no luck germinating spores on previous plates so I just left the drop of solution in the corner hoping two spores meet up. Just for kicks I will be trying to isolate a few of these sites and transfer them onto new plates. Of course I can not find any stainless steel wire (it is in a box here somewhere) but then I had a thought. I could use the end of a guitar string as one.

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wavyedge said:

I know gentamicin survives autoclaving temps, so you add when you mix. Best to look up the guidance on whatever you add individually.

Antibiotics can have a real effect on mycelial growth and health, so best to only use for trying to clean up a transfer, and drop onto normal media for growth and expansion.

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anders said:

Gentamicin is gram-negative, and evidently hasn't worked, which imples that the bacteria we are concerned with is gram-positive.

Chloramphenicol is gram-positive, and may do it.

Trichoderma (the green) is a mold, not bacterial.

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RoscoeReturns said:

Gentamicin is heat stable and will survive autoclaving just fine. You can add that to your broth before you sterilize.

Chloramphenicol is NOT heat stable. After you autoclave your media you must wait until it cools before adding chloramphenicol. It must be added aseptically to your liquid media just before you pour. Chloramphenicol is also degraded by hydrolysis in an aqueous vehicle. Use those plates soon after you pour them, they don’t retain activity for very long. Refrigeration can slow that process.

Malt extract agar already inhibits the growth of most bacterial species due to its acidic pH. Key words being inhibits growth, not kills off. I’m currently experimenting with using tryptic soy agar media to see if this growth inhibition is causing “false negatives.” If bacteria is present yet not able to grow am I seeing a clean plate that is not really clean?

Having said all that, I have used antibiotic agar in the past to clean up very dirty cultures that I couldn’t otherwise replace. Other than that very rare (for me) instance, I don’t see the utility of it in cultivation. I’ve had equal success using cabin sequestering, so I don’t know that I’ll go through the added steps of making antibiotic agar again.

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anders said:

H2O2 even at 12% is not the best.  But I am not supposed to say anything negative.

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Negative away my friend, I'm all for the constructive criticism and feedback

I am interested to hear what you have to say

Puffin said:

anders said:

H2O2 even at 12% is not the best.  But I am not supposed to say anything negative.

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Negative away my friend, I'm all for the constructive criticism and feedback

I am interested to hear what you have to say

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anders said:

I have received my first and what I presume to be my only warning in backchannel, for impugning a Mod who I did not know was a Mod, after he insulted me.  I do not need this kind of treatment and do not tolerate this temerity at my age and education level.

There is a better option than h2o2 or denatured alcohol.  You can figure out if you look around. (not here)

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RoscoeReturns said:

If bacteria is present yet not able to grow am I seeing a clean plate that is not really clean?

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wavyedge said:

RoscoeReturns said:

If bacteria is present yet not able to grow am I seeing a clean plate that is not really clean?

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Fuck me. Good point. Cooking me some potatoes for dinner tonight.

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Puffin said:

I did have another idea about isolating monokaryons which is to filter the spore solution through a 20 μm filter prior to cell counting and dilution with the aim of eliminating the majority of the clumps and increasing the single spore ratio.

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PhenoDreamers said:

Let's leave h2o2 out of this. I've at least proven it's definitely effective. It's certainly better than iso. However, people always want to use it in manners that are impractical. I don't use iso at all. Only h2o2.

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