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growbit
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Help! First time agar - is it supposed to look like this?
#23187230 - 05/04/16 01:11 AM (8 years, 8 months ago) |
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I put a single healthy colonized grain onto agar about 5 days ago, and so far the myc hasn't crept into the Agar yet. I'm thinking I may have made the nutrients too dense - I used potato flakes and honey.
The trouble is the myc has grown into a large puffy ball on top of the agar. I want to prepare it for isolation but there are no obvious sectors as it hasn't crept into the agar yet.
The view from the side describes the scenario:
 I want to take a small piece of this and make a liquid culture, and also transfer some sectors out to make grain masters for fruiting. How much longer should I wait?
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mushpunx
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Re: Help! First time agar - is it supposed to look like this? [Re: growbit]
#23187269 - 05/04/16 01:38 AM (8 years, 8 months ago) |
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Its just puffing up, it'll spread out onto the agar just give it time.
As soon as it does youre going to want to take a transfer to another plate, to make sure youre working wstarting lean culture before you do anything with it.
Can you take a photo of the plate from the top ? I cant tell if Im looking at bacteria or not.
You arent going to see sectoring on this plate. It usually takes *at least* 10 transfers before you see any obvious sectoring. It might take a couple less for you since you are starting with a grain.
If you are looking to isolate strains, you will need to transfer probably ten times or more. Then you will isolate each sector to a new plate. You will then have to keep transferring from each of those plates untill you no longer see any sectoring... then label and test every culture... keeping the good ones.
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growbit
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Re: Help! First time agar - is it supposed to look like this? [Re: mushpunx]
#23187546 - 05/04/16 05:28 AM (8 years, 8 months ago) |
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Here's a picture from the top. The discoloration you see might not be bacteria, but water dripping from the top of the dish from condensation since I used a no-pour tek. Your thoughts?
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Inocuole
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Re: Help! First time agar - is it supposed to look like this? [Re: growbit]
#23187598 - 05/04/16 06:08 AM (8 years, 8 months ago) |
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No, it's bacteria. It looks like a lighter colored liquid but that's how it looks.
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growbit
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Re: Help! First time agar - is it supposed to look like this? [Re: Inocuole]
#23187641 - 05/04/16 06:33 AM (8 years, 8 months ago) |
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What would the best strategy be here to salvage the mycelium?
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Josex
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Re: Help! First time agar - is it supposed to look like this? [Re: growbit]
#23187667 - 05/04/16 06:45 AM (8 years, 8 months ago) |
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Quote:
growbit said: What would the best strategy be here to salvage the mycelium?
You should take a wedge as soon as you can and as far away from contams as possible and transfer it to a blank agar plate. A small triangular wedge will do the job.
RR has incredibly usefull videos about agar and sterile technique, I'm sure it will help you a lot. You may want to take a look at this as well.
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growbit
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Re: Help! First time agar - is it supposed to look like this? [Re: Josex]
#23187739 - 05/04/16 07:30 AM (8 years, 8 months ago) |
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Quote:
Josex said:
Quote:
growbit said: What would the best strategy be here to salvage the mycelium?
You should take a wedge as soon as you can and as far away from contams as possible and transfer it to a blank agar plate. A small triangular wedge will do the job.
RR has incredibly usefull videos about agar and sterile technique, I'm sure it will help you a lot. You may want to take a look at this as well.
Rockstar, thank you.
Quick question, from the above pictures how sure are you that the white part actually is mycelium? It's very white but its so puffy its unlike any agar pictures I've personally seen. Is there a chance that there is hidden mold riding along the myc making it look like that?
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mushpunx
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Re: Help! First time agar - is it supposed to look like this? [Re: Inocuole]
#23187761 - 05/04/16 07:40 AM (8 years, 8 months ago) |
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Quote:
Inocuole said: No, it's bacteria. It looks like a lighter colored liquid but that's how it looks.
Yea I see bactetia blobs... you can see it looks like slime/boogers at Five O'clock is real obvious, but it looks like a layer of sneaky bacteria over the whole left side of the plate. You could try to take a transfer from up around 3 oclock or so (best I can tell from the pic).
Are you working in a still air box?
The myc looks normal to me
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growbit
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Re: Help! First time agar - is it supposed to look like this? [Re: mushpunx]
#23187788 - 05/04/16 07:52 AM (8 years, 8 months ago) |
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Thanks for clarifying, glad I at least have some good myc in there.
I was working in a still air box, but I opened a jar of contaminated WBS to save some myc inside, and its obvious some of those contams have gone into the agar too. I'll take a 3 o' clock sector out in the next day or two once the myc has grown onto the agar a bit more. Or should I try do a transfer now?
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mushpunx
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Re: Help! First time agar - is it supposed to look like this? [Re: growbit]
#23187801 - 05/04/16 07:57 AM (8 years, 8 months ago) |
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Quote:
growbit said: Thanks for clarifying, glad I at least have some good myc in. here.
I was working in a still air box, but I opened a jar of contaminated WBS to save some myc inside, and its obvious some of those contams have gone into the agar too. I'll take a 3 o' clock sector out in the next day or two once the myc has grown onto the agar a bit more. Or should I try do a transfer now?
You should take it now before its too late. You only need a peice But honestly, if the jar the grain came from was contaminated I wouldn't even bother with that culture. Start a new one.
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