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Yoyoma
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Registered: 07/24/18
Posts: 34
Last seen: 3 years, 5 months
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Is this normal growth??
#26802565 - 07/04/20 12:44 AM (3 years, 6 months ago) |
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Help me identify if this is normal growth for these plates so far. I'm a noob doing agar for the first time. I will list what I did and my mistakes and the photos following. Please let me know if anything is the results are my mistakes.
-June 28 innc. (5 days) -Used 4 pre-poured agar plates -Used 2 different spore syringes (B+ plates 1/2, Golden T plates 3/4) -SAB -Did not shake the syringes prior -Flame sterilized needle only before plate 1 and 3 -Did not cool the needle in agar before squirting -Squirted multiple drops because I was nervous -Did NOT go back and streak with loop
edit: ignore the black markings (reflection)
PLATE 1 seems like nothing is going on



PLATE 2 seems like Mycelium growth



PLATE 3 not sure what's going on here



PLATE 4 also not sure


Edited by Yoyoma (07/04/20 12:58 AM)
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Gastronomicus
3-0-G



Registered: 03/31/05
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Re: Is this normal growth?? [Re: Yoyoma]
#26802613 - 07/04/20 01:12 AM (3 years, 6 months ago) |
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Plate 1 is contaminated unless those specks are sediment in the agar, plate 3 and 4 are totally fucked.
Plate 2 looks like myc but it's also super wispy and disorganized and might be mold. Take a transfer to a clean plate and wait and see.
-------------------- Make my Funk the P Funk, I wants to get Funked up
LAGM2024
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Yoyoma
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Registered: 07/24/18
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Last seen: 3 years, 5 months
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pretty sure it's specs in the agar on plate 1
should I go ahead and transfer plate 2 already? Any small edge??
what on earth is going on with 3 and 4??
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SoupyGeorge
Hazy Daze since First Grade



Registered: 05/25/20
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Loc: Southern US of 'Merica
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Re: Is this normal growth?? [Re: Yoyoma]
#26802620 - 07/04/20 01:19 AM (3 years, 6 months ago) |
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Spores ball up in the syringes. Shaking is a must to break the spore clusters up. It's possible your first plate didn't get any spores. Wait a few days to make sure though.
I did not cool my needle either and my plates colonized just fine. Your second plate indeed looks like Mycelium.
The two last plates are both contaminated with bacteria, that's the puddle like substance you see. Possibly from the syringe itself considering they are the only two with that contamination.
It is best to only use one single drop from a very well shaken syringe and either streak it with an inoculation loop or let it roll in a straight line from on side of the dish to the other and set it down not disturbing it for a while. Both methods have worked for me.
-------------------- Mellow heads will prevail. You look like everyone else because the T.V. tells you to. I look like everyone else so that i don't stand out to the man. We are not the same.
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Yoyoma
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Registered: 07/24/18
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Last seen: 3 years, 5 months
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Thank you for the info!
Yes, I was nervous so I did forget a few steps. I did not realize I wouldn't be able to see the drops in the agar. I also only had 4 agar plates. I won't be able to make more either until a week from now, so any more inoculations or transfer of plate 2 will have to wait.
Disappointed and a little surprised about plate 3/4 as I thought my technique had improved from the previous plates lol. But as you stated it must have been from the syringe. Is this just a matter of getting a good single drop spread?
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redhandmat
Dude


Registered: 05/09/19
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Re: Is this normal growth?? [Re: Yoyoma]
#26802668 - 07/04/20 01:53 AM (3 years, 6 months ago) |
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From the plate that looks like myc, you should take transfers (T1) as soon as you can. very small from the best looking area.
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Yoyoma
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Last seen: 3 years, 5 months
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Re: Is this normal growth?? [Re: redhandmat]
#26802684 - 07/04/20 02:02 AM (3 years, 6 months ago) |
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Would I ever take it from the middle or only from the edges?
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redhandmat
Dude


Registered: 05/09/19
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Re: Is this normal growth?? [Re: Yoyoma]
#26802696 - 07/04/20 02:20 AM (3 years, 6 months ago) |
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Quote:
Yoyoma said: Would I ever take it from the middle or only from the edges?
The edge is where you want to take your transfers.
In your future plates try to get clear agar with no sediments so that you are truly in the know of what is what. Check teks on how to get it like that. What I personally do is I have clear LME in powder form, add it with the agar (right amount of both ofc), add the right amount of water (I like adding it boiling hot but that's not necessary probably). Then I use a mixer or manually mix it really well while its boiling. The results are clear agar plates that you can see through (like glass).
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