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OfflineFriskydingo
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Using agar
    #23798887 - 11/03/16 07:47 PM (7 years, 3 months ago)

Hey all. Been lurking the site for a bit now, gleaned tons of useful information from you all. Many thanks.
I've had at least ten successful mono tub grows now. I tried doing agar and it really didn't seem to work out. Was growing some but seemed to just stall out. I also noticed looking at the petri dish from the bottom the grow site was very dark instead of white like the top. I understand it grows on top but the darkness on the bottom concerned me. I wound up tossing them so I can refine things a bit. Any input on the dark bottom would be appreciated.

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Offlinenumnum59
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Re: Using agar [Re: Friskydingo]
    #23798893 - 11/03/16 07:49 PM (7 years, 3 months ago)

Use foodcoloring? How thick was the agar?

Sounds like possible contamination but without pictures its hard to tell.

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OfflineFriskydingo
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Re: Using agar [Re: numnum59]
    #23798990 - 11/03/16 08:19 PM (7 years, 3 months ago)

I did not use food coloring. The agar powder I used had a brownish tint to it already. I did see some spots that were obviously contams. I did so many things wrong when I did them but it was a learning process for me. I tried posting a picture on some questions about my tubs but they were the wrong format or something and I gave up. Time answered my questions for me anyway.
Oh, as for the thickness of the agar it was just enough to cover the bottom of the dish.

Edited by Friskydingo (11/03/16 08:22 PM)

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Offlinenumnum59
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Re: Using agar [Re: Friskydingo]
    #23799016 - 11/03/16 08:27 PM (7 years, 3 months ago)

Well it is a learning experience for sure

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OfflineKryptos
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Re: Using agar [Re: numnum59]
    #23799213 - 11/03/16 09:53 PM (7 years, 3 months ago)

I use charcoal powder (~.5g/L) instead of food coloring for my agar. Makes the white mycelium very visible against the black agar.

I also try to avoid tossing agar dishes whenever possible. I've managed to pull clean isolates from some seriously nasty plates before. Sure, it usually takes 2-3 transfers, but it feels really good when you get that pure white isolate from a rainbow plate. Just be careful you don't contaminate *everything*. It's also good practice for when you're trying to isolate a wild mushroom and every sample that you have is trash/bacterial/partly digested.

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OfflineFriskydingo
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Re: Using agar [Re: Kryptos]
    #23799556 - 11/04/16 12:39 AM (7 years, 3 months ago)

I pretty much tossed them because of condensation made them difficult to look at what was going on inside and they stalled out. After learning more about technique to avoid the condensation I just figured I'd start over. I have plenty of agar and dishes. How long should it take for the sample to grow out? I did a few grains from jars, a few clones and some from ms syringe. The clones were a real problem getting off the blade so weren't exactly centered and shaking the dishes to try and center left a interesting trail across the agar. The MS was growing but appeared very thin and faint. They had obvious contam on the agar. The grains seemed to be growing the best. I'm pretty stoked about doing it, I think it's fascinating as fuck, playing God with this shit lol
I ordered a Halloween special from a vendor and it will have some Pans in it so I'm looking forward to working with them.

Edited by Friskydingo (11/04/16 01:32 PM)

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Offlinenumnum59
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Re: Using agar [Re: Kryptos]
    #23799833 - 11/04/16 06:39 AM (7 years, 3 months ago)

Quote:

Kryptos said:
I use charcoal powder (~.5g/L) instead of food coloring for my agar. Makes the white mycelium very visible against the black agar.

I also try to avoid tossing agar dishes whenever possible. I've managed to pull clean isolates from some seriously nasty plates before. Sure, it usually takes 2-3 transfers, but it feels really good when you get that pure white isolate from a rainbow plate. Just be careful you don't contaminate *everything*. It's also good practice for when you're trying to isolate a wild mushroom and every sample that you have is trash/bacterial/partly digested.




Seems like it would be hard to spot contamination with a black background.

I agree with trying to get clean pulls from each plate its the best way to learn.

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