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Zombie-ant
Mushroom Ninja in training



Registered: 09/02/19
Posts: 549
Loc: UK
Last seen: 16 days, 13 hours
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Agar growth?
#26501729 - 02/24/20 03:10 PM (3 years, 10 months ago) |
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Or not or too early, contam?
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iwh678
Budget Mycologist



Registered: 01/22/14
Posts: 396
Loc: USA
Last seen: 1 year, 6 months
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Those satellite colonies are all bacteria. A little hard to tell on the center one from this pic.
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Zombie-ant
Mushroom Ninja in training



Registered: 09/02/19
Posts: 549
Loc: UK
Last seen: 16 days, 13 hours
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Re: Agar growth? [Re: iwh678]
#26501784 - 02/24/20 03:51 PM (3 years, 10 months ago) |
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Is this image any better
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Neowynd8

Registered: 06/12/16
Posts: 277
Last seen: 7 months, 1 day
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Trash
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iwh678
Budget Mycologist



Registered: 01/22/14
Posts: 396
Loc: USA
Last seen: 1 year, 6 months
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Re: Agar growth? [Re: Neowynd8]
#26501967 - 02/24/20 05:24 PM (3 years, 10 months ago) |
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That also looks bacterial.
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mushboy
modboy



Registered: 04/24/05
Posts: 32,274
Loc: where?
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Re: Agar growth? [Re: iwh678]
#26501969 - 02/24/20 05:25 PM (3 years, 10 months ago) |
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Ya that's jacked up.
What kind of scalpel do you use?
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Sockadin



Registered: 01/03/10
Posts: 7,244
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Re: Agar growth? [Re: mushboy]
#26502097 - 02/24/20 06:45 PM (3 years, 10 months ago) |
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Plastic knife.
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mushboy
modboy



Registered: 04/24/05
Posts: 32,274
Loc: where?
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Re: Agar growth? [Re: Sockadin]
#26502105 - 02/24/20 06:49 PM (3 years, 10 months ago) |
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Hey ...wait a minute! your not the OP!?
Only asking cause all that bacteria could come from the scalpel handle. That's why I went to an all metal one.
Flame sterilizing plastic is kinda tricky
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Sockadin



Registered: 01/03/10
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Re: Agar growth? [Re: mushboy]
#26502145 - 02/24/20 07:10 PM (3 years, 10 months ago) |
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I'm sorry.... I posted that one and thought, hmm maybe that's to harsh.
Edited by Sockadin (02/24/20 07:11 PM)
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Zombie-ant
Mushroom Ninja in training



Registered: 09/02/19
Posts: 549
Loc: UK
Last seen: 16 days, 13 hours
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Re: Agar growth? [Re: Sockadin]
#26502462 - 02/25/20 12:02 AM (3 years, 10 months ago) |
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Quote:
Sockadin said: I'm sorry.... I posted that one and thought, hmm maybe that's to harsh.
🤣
Plastic Knife.... I wouldn’t put it past me.
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Zombie-ant
Mushroom Ninja in training



Registered: 09/02/19
Posts: 549
Loc: UK
Last seen: 16 days, 13 hours
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Re: Agar growth? [Re: mushboy]
#26502463 - 02/25/20 12:06 AM (3 years, 10 months ago) |
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Quote:
mushboy said: Hey ...wait a minute! your not the OP!?
Only asking cause all that bacteria could come from the scalpel handle. That's why I went to an all metal one.
Flame sterilizing plastic is kinda tricky
I used an innoc loop. Following bod‘s agar vid.
I used a candle to flame, maybe not hot enough am waiting on a mini flamer from eBay.
I have 99% isopropyl, I diluted it slightly, as bods vid says a lower percentage (vid didn’t mention of 99% was higher, that was probably dumb) Which of the above or is all the above dumbass? 🙄
I have a metal scalpel handle (and metal blade, not plastic) and metal inoculation loops. I can use 99% isopropyl to dip before flaming right, or does the percentage have to be lower?
I have more glass dishes, that I will be dry sterilising as some point.
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Edited by Zombie-ant (02/25/20 12:16 AM)
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Neowynd8

Registered: 06/12/16
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What you dip it in is irrelevant as long as you flame sterilize properly. I'd say a candle is good enough. Make sure the tool gets red hot.
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Zombie-ant
Mushroom Ninja in training



Registered: 09/02/19
Posts: 549
Loc: UK
Last seen: 16 days, 13 hours
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Re: Agar growth? [Re: Neowynd8]
#26502594 - 02/25/20 05:23 AM (3 years, 10 months ago) |
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Quote:
Neowynd8 said: What you dip it in is irrelevant as long as you flame sterilize properly. I'd say a candle is good enough. Make sure the tool gets red hot.
I did make sure it was red hot. Anyway I have loads more spore prints to work through. This one was thin, not sure that has anything to do with. At least I can grow bacteria 🦠💩☹️😂
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MrBovineJoni
Glip-Glop


Registered: 11/28/19
Posts: 264
Last seen: 13 days, 13 hours
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Quote:
Zombie-ant said:
Quote:
Neowynd8 said: What you dip it in is irrelevant as long as you flame sterilize properly. I'd say a candle is good enough. Make sure the tool gets red hot.
I did make sure it was red hot. Anyway I have loads more spore prints to work through. This one was thin, not sure that has anything to do with. At least I can grow bacteria 🦠💩☹️😂
One good print can be used to inoculate 10s of plates so I would not worry about it too much (as there are millions of spores per print and you only need a few thousands for a few germination points - if the spores are not very old), just typically bacteria shows up before spores have germinated and started to grow.
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Zombie-ant
Mushroom Ninja in training



Registered: 09/02/19
Posts: 549
Loc: UK
Last seen: 16 days, 13 hours
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Quote:
typically bacteria shows up before spores have germinated and started to grow.
Hence why agar is good, as bacteria starts first so it can be removed, right?
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iwh678
Budget Mycologist



Registered: 01/22/14
Posts: 396
Loc: USA
Last seen: 1 year, 6 months
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No, do not remove contams from your dish. You want to transfer clean mycelium away from the contamination and clean it up through more transfers.
You can leave these to see if mycelium grows. They're not going to hurt anything. In the mean time I would get some new dishes started.
Is it possible your issue is from the pour itself? Have you left any blank plates to the side for a while to see if they stay clean?
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Zombie-ant
Mushroom Ninja in training



Registered: 09/02/19
Posts: 549
Loc: UK
Last seen: 16 days, 13 hours
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Re: Agar growth? [Re: iwh678]
#26502617 - 02/25/20 05:53 AM (3 years, 10 months ago) |
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These innocs are only four days old, I should have said. So still early days no? Other plates are available, I am reheat sterilising some glass plates I have, I have ordered an oven thermometer, as I think my oven may be shit. The print was about 6 months old.
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Zombie-ant
Mushroom Ninja in training



Registered: 09/02/19
Posts: 549
Loc: UK
Last seen: 16 days, 13 hours
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Re: Agar growth? [Re: iwh678]
#26502618 - 02/25/20 05:56 AM (3 years, 10 months ago) |
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Quote:
iwh678 said: No, do not remove contams from your dish. You want to transfer clean mycelium away from the contamination and clean it up through more transfers.
You can leave these to see if mycelium grows. They're not going to hurt anything.
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Zombie-ant
Mushroom Ninja in training



Registered: 09/02/19
Posts: 549
Loc: UK
Last seen: 16 days, 13 hours
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Re: Agar growth? [Re: iwh678]
#26502626 - 02/25/20 06:14 AM (3 years, 10 months ago) |
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Quote:
iwh678 said:
Is it possible your issue is from the pour itself?
The agar was sterilised, it set then I diluted it to make make the mixture go further, I sterilised it again though after. I covered the lid, but opened it a couple of timeS while cooling, but not much 10ish secs. a couple of times while it was streaming hot. I wore a Tyvek suit, face mask, Sanitised gloves Using an SAB too.
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Edited by Zombie-ant (02/25/20 06:16 AM)
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iwh678
Budget Mycologist



Registered: 01/22/14
Posts: 396
Loc: USA
Last seen: 1 year, 6 months
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Quote:
Zombie-ant said: These innocs are only four days old, I should have said. So still early days no? Other plates are available, I am reheat sterilising some glass plates I have, I have ordered an oven thermometer, as I think my oven may be shit. The print was about 6 months old.
What are you using an oven for?
Quote:
Zombie-ant said:
Quote:
iwh678 said:
Is it possible your issue is from the pour itself?
The agar was sterilised, it set then I diluted it to make make the mixture go further, I sterilised it again though after. I covered the lid, but opened it a couple of timeS while cooling, but not much 10ish secs. a couple of times while it was streaming hot. I wore a Tyvek suit, face mask, Sanitised gloves Using an SAB too.
Where did you open it up?
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Zombie-ant
Mushroom Ninja in training



Registered: 09/02/19
Posts: 549
Loc: UK
Last seen: 16 days, 13 hours
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Re: Agar growth? [Re: iwh678]
#26502652 - 02/25/20 06:44 AM (3 years, 10 months ago) |
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Quote:
What are you using an oven for?
Dry sterilising glass plates wrapped in foil.
Where did you open it up?
When I put it in a cooling bath...
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MrBovineJoni
Glip-Glop


Registered: 11/28/19
Posts: 264
Last seen: 13 days, 13 hours
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Quote:
Zombie-ant said:
Quote:
iwh678 said:
Is it possible your issue is from the pour itself?
The agar was sterilised, it set then I diluted it to make make the mixture go further, I sterilised it again though after. I covered the lid, but opened it a couple of timeS while cooling, but not much 10ish secs. a couple of times while it was streaming hot. I wore a Tyvek suit, face mask, Sanitised gloves Using an SAB too.
Ideally you don't want the receiver dish petri lid to be open for longer than a second or two when working in SAB. Every second increases the chance of contamination - get spores on the sterilised loop; open the lid, swipe with the loop in the chosen pattern and carefully close the lid as soon as possible (while still doing steady motions inside SAB).
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Zombie-ant
Mushroom Ninja in training



Registered: 09/02/19
Posts: 549
Loc: UK
Last seen: 16 days, 13 hours
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Quote:
MrBovineJoni said:
Ideally you don't want the receiver dish petri lid to be open for longer than a second or two when working in SAB. Every second increases the chance of contamination - get spores on the sterilised loop; open the lid, swipe with the loop in the chosen pattern and carefully close the lid as soon as possible (while still doing steady motions inside SAB).
I will have to practice my Mushroom Ninja Skills
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