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OfflineSpiritwithin
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Look at my plates... what do you see?
    #23391276 - 06/28/16 02:36 PM (7 years, 6 months ago)

Fellow mycologists,

I've done some agar work... these are all second transfers from GT MS.
Now I see some sectoring in the inner part (1st and 2nd picture) but should I not take samples from the outer rim for my next transfer?

I'm pretty positive that this is not mold, but I  could be wrong of course...

Any advice would be appreciated.

Thx



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OfflineHybridprX
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Re: Look at my plates... what do you see? [Re: Spiritwithin]
    #23391294 - 06/28/16 02:41 PM (7 years, 6 months ago)

Take them from the thickest/fastest outer growth. I can see a few sectors worthy of transfer.


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OfflineKenetic
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Re: Look at my plates... what do you see? [Re: Spiritwithin]
    #23391305 - 06/28/16 02:44 PM (7 years, 6 months ago)

Ha I see the fairest maiden of them all....in about 6 or 7 transfers....:smile:

Edit: looks like your plates are a litte overrun.


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Edited by Kenetic (06/28/16 02:46 PM)


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OfflineSpiritwithin
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Re: Look at my plates... what do you see? [Re: HybridprX]
    #23391308 - 06/28/16 02:46 PM (7 years, 6 months ago)

Quote:

HybridprX said:
Take them from the thickest/fastest outer growth. I can see a few sectors worthy of transfer.




Thanks for answering. I want to make three new plates... Would you be so kind as to tell me which sectors you would choose?
Maybe in a "2nd picture 8 o'clock" way...
I would very much appreciate it!


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OfflineSpiritwithin
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Re: Look at my plates... what do you see? [Re: Kenetic]
    #23391331 - 06/28/16 03:01 PM (7 years, 6 months ago)

Quote:

kenetic said:
Ha I see the fairest maiden of them all....in about 6 or 7 transfers....:smile:

Edit: looks like your plates are a litte overrun.




Tomorrow I will transfer to new plates :wink:... If I figure out from where to take the samples until then. Maybe it won't make such a big difference...
I think when the temperature droped quite a bit a few days ago, the mycelium suddenly grew thicker. Could that be?


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OfflineKenetic
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Re: Look at my plates... what do you see? [Re: Spiritwithin]
    #23391361 - 06/28/16 03:19 PM (7 years, 6 months ago)

Your best bet it to take several samples from different, similarly aggressive growth areas and branch off from there, simply because you seem to be trying to isolate a random strain from multispore.  Dont just get one isolate becase it could be anything.  You are gonna need a lot of plates (40?) To isolate a few strains and then you need to test each one seperately. 
But to answer your immediate question I would try the first pic at 8 (am or pm, your choice), first pic at 10, and second pic at noon, for starters.
Next time try isolating a clone, you have way less genetics to work with.


--------------------
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Re: Look at my plates... what do you see? [Re: Kenetic]
    #23391369 - 06/28/16 03:24 PM (7 years, 6 months ago)

Isolating a clone will still give you a clone. Getting a true isolate is a fun and exciting process. For me anyway.


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OfflineSpiritwithin
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Re: Look at my plates... what do you see? [Re: Kenetic]
    #23391407 - 06/28/16 03:39 PM (7 years, 6 months ago)

Quote:

kenetic said:
Your best bet it to take several samples from different, similarly aggressive growth areas and branch off from there, simply because you seem to be trying to isolate a random strain from multispore.  Dont just get one isolate becase it could be anything.  You are gonna need a lot of plates (40?) To isolate a few strains and then you need to test each one seperately. 
But to answer your immediate question I would try the first pic at 8 (am or pm, your choice), first pic at 10, and second pic at noon, for starters.
Next time try isolating a clone, you have way less genetics to work with.




Ok. Thank you very much!

Now about the clone: I thought about that, but, If I clone 1 mushroom I cannot test its potency since 1 shroom is way to less weight to trip. So even though it may look nice and healthy its still a matter of sheer luck, is it not?


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OfflineKenetic
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Re: Look at my plates... what do you see? [Re: Spiritwithin]
    #23391467 - 06/28/16 04:03 PM (7 years, 6 months ago)

No thats why you test them.  Grow the isolates out pf style or something and look for desirable traits.
You dont have to eat just one shroom test it out.  Measure out a gram or something.
Im too lazy for all of this.  I just get a clone tissue sample and grow it out on a half inch of rye and g2g. 
In your case just keep using agar, but try it sometime with a clone.


--------------------
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                                                :cookiemonster::elmo:



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Invisibledankington
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Re: Look at my plates... what do you see? [Re: Kenetic]
    #23391531 - 06/28/16 04:24 PM (7 years, 6 months ago)

third plate sure looks pretty nice to me!


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Re: Look at my plates... what do you see? [Re: dankington]
    #23392788 - 06/29/16 12:34 AM (7 years, 6 months ago)

Really? I think I'd prefer to with 1 and 2. Don't like wispy cultures all that much.


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Offlinespacechildo
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Re: Look at my plates... what do you see? [Re: Spiritwithin]
    #23393232 - 06/29/16 07:02 AM (7 years, 6 months ago)

still looks fucked to me :shrug:

making a new thread doesnt change the fact, maybe it can give you the answer you're looking for tho :goodluck:


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Re: Look at my plates... what do you see? [Re: spacechildo]
    #23393246 - 06/29/16 07:11 AM (7 years, 6 months ago)

im fairly new to agar, and super new to isolating sectors and what not, but if it were me, i would take from plate 1 at exact 6o'clock, where the myc sorta juts out a tiny bit. i would also try to get right around 1:45 and 12:30, where they jut out.

and for plate 2 i would try to get right around 3:00-3:15, and then around 11:00 and 2. for all those, its where you can see it jut out a little, and sorta looks stronger.

as for plate 3, idk haha..

again, im super new to this, but thats probably what i would choose.


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Re: Look at my plates... what do you see? [Re: mupetmower]
    #23393263 - 06/29/16 07:19 AM (7 years, 6 months ago)

just look at all the separating lines, definitely not clean


cube myc and cube myc grow together, cube myc and foreign contam myc separates like this.


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Re: Look at my plates... what do you see? [Re: spacechildo]
    #23393269 - 06/29/16 07:21 AM (7 years, 6 months ago)

it is still salvageable, right? idk haha. like you could still take what looks like it's growing ok and transfer and clean it up, right?


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Offlinespacechildo
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Re: Look at my plates... what do you see? [Re: mupetmower]
    #23393275 - 06/29/16 07:24 AM (7 years, 6 months ago)

and hope mold isnt intertwined in your transfer? sure.
but why hope when you can start over and do it right?

Starting off a grow from clearly contamed plates isn't something I'd recommend.


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Re: Look at my plates... what do you see? [Re: spacechildo]
    #23393281 - 06/29/16 07:26 AM (7 years, 6 months ago)

oh. i just thought that was something that people did regularly? taking transfers from clearly contam'd plates and cleaning it up with multiple transfers, to be able to use.

Not being sarcastic or anything, i just really thought that was one of the main purposes of using agar.


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-I wanna feel the change consume me, feel the outside turning in. I wanna feel the metamorphosis and cleansing I've endured within.


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Re: Look at my plates... what do you see? [Re: mupetmower]
    #23393287 - 06/29/16 07:30 AM (7 years, 6 months ago)

yeah, but when the mold is intertwined in the myc how do you separate them?
if you get some satelites or mold growing on 1 side just transfer away. I doubt OP will be able to get the mold away from his myc since its all mingled together.


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Re: Look at my plates... what do you see? [Re: spacechildo]
    #23393290 - 06/29/16 07:30 AM (7 years, 6 months ago)

ohh.. hmm.. ive still got a lot to learn, then.

so, in this case, you just toss the first two plates and transfer from 3?


--------------------
-The wise man never stops seeking knowledge.


-I wanna feel the change consume me, feel the outside turning in. I wanna feel the metamorphosis and cleansing I've endured within.


Edited by mupetmower (06/29/16 07:31 AM)


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Offlinespacechildo
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Re: Look at my plates... what do you see? [Re: mupetmower]
    #23393295 - 06/29/16 07:33 AM (7 years, 6 months ago)

Quote:

mupetmower said:
like you could still take what looks like it's growing ok and transfer and clean it up, right?




nothing looks like its growing OK to me, its all got mold mixed in.
maybe thats where our confusion came from.

water is 1/3 oxygen but breathing under water is still pretty impossible since you cant single out the oxygen molecules (or cube myc in this scenario) out of the water (contams)


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Re: Look at my plates... what do you see? [Re: spacechildo]
    #23393298 - 06/29/16 07:36 AM (7 years, 6 months ago)

I've had clean cultures look like that when the mycelium has a hard time staying above the surface. The thicker growth is where the mycelium finally manages to break the surface. It is still a good idea to make transfers before using that plate as an inoculant though, having a uniform plate brings more peace of mind than a patched plate like that. Doesn't mean it's dirty, just means don't trust it just yet.


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Re: Look at my plates... what do you see? [Re: Supalemonhaze]
    #23393303 - 06/29/16 07:38 AM (7 years, 6 months ago)

Quote:

Supalemonhaze said:
I've had clean cultures look like that when the mycelium has a hard time staying above the surface. The thicker growth is where the mycelium finally manages to break the surface.




say what? surface of the agar? why would myc have a hard time on agar?
I've never had growth like this turn out clean, its separating because its meeting an enemy.


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Re: Look at my plates... what do you see? [Re: spacechildo]
    #23393304 - 06/29/16 07:39 AM (7 years, 6 months ago)

heh great analogy.

and yeah, i didnt mean to use it as inoculant, but just to transfer from it, still, to see if it can be saved. but, i guess if its only a second transfer from MS, and you have more spores, then maybe it would be easier to just start over.


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-I wanna feel the change consume me, feel the outside turning in. I wanna feel the metamorphosis and cleansing I've endured within.


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Re: Look at my plates... what do you see? [Re: spacechildo]
    #23393322 - 06/29/16 07:47 AM (7 years, 6 months ago)

Quote:

spacechildo said:
Quote:

Supalemonhaze said:
I've had clean cultures look like that when the mycelium has a hard time staying above the surface. The thicker growth is where the mycelium finally manages to break the surface.




say what? surface of the agar? why would myc have a hard time on agar?





:shrug: Beats me, always only happened to me when I was using GWA though so I'm guessing too much nutes or something. Once I transferred back on LMEA it grew normally. Meshed in mold usually grows in "patches" IME but I don't have much experience with those.


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Re: Look at my plates... what do you see? [Re: Supalemonhaze]
    #23393338 - 06/29/16 07:55 AM (7 years, 6 months ago)

but what do you mean when you say break the surface? as if the myc was growing on the underside of the agar trying to break through it?


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Re: Look at my plates... what do you see? [Re: spacechildo]
    #23393345 - 06/29/16 07:58 AM (7 years, 6 months ago)

Yeah, it would seem like it's growing under the surface. You can still see the mycelium above the surface if you use a light but it doesn't grow thick until it manages to surface properly. Once it does, it always looked like the first two pics in the OP, where light growth turns into thich growth and then settles down for the rest of the plate. When this happened to me, I could see the agar being pushed up where the mycelium is trying to colonize through it.


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Re: Look at my plates... what do you see? [Re: Supalemonhaze]
    #23393354 - 06/29/16 08:02 AM (7 years, 6 months ago)

you're talking about a hot pour or sandwich tek right?
you havent held a light up to OPs plates?
I'm really confused now :lol:

I think OP said this was a transfer or 2 away from original plate in his other thread..


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Re: Look at my plates... what do you see? [Re: Supalemonhaze]
    #23393358 - 06/29/16 08:04 AM (7 years, 6 months ago)

i actually have two clone plates that seemed to have done the same thing. they had very thin, wispy looking growth, that even seemed to be below the surface, and then all the sudden, it started growing from the top, and got thick, and healthier looking. but with these, thee growth did it in a perfect circle, and not in weird lines like OP's.

I mean, im sure it couldve easily been bacteria or mold that caused this to happen, im still too new to properly identify what is actually going on with it. but i transferred from it, so we will see what happens.

Oh, and yeah, space, OP said that these plates are second transfer from original i think.


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-I wanna feel the change consume me, feel the outside turning in. I wanna feel the metamorphosis and cleansing I've endured within.


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Re: Look at my plates... what do you see? [Re: spacechildo]
    #23393362 - 06/29/16 08:06 AM (7 years, 6 months ago)

No lol. I'm talking about regular transfers made on GWA which appeared very similiar to OP's and turned out to be clean.


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Re: Look at my plates... what do you see? [Re: Supalemonhaze]
    #23393370 - 06/29/16 08:11 AM (7 years, 6 months ago)

This is what i was talking about:



thats a plate made from a tissue sample from an Ecuadorian fruit that grew in another plate.


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-The wise man never stops seeking knowledge.


-I wanna feel the change consume me, feel the outside turning in. I wanna feel the metamorphosis and cleansing I've endured within.


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Re: Look at my plates... what do you see? [Re: Supalemonhaze]
    #23393378 - 06/29/16 08:15 AM (7 years, 6 months ago)

Quote:

Supalemonhaze said:
No lol. I'm talking about regular transfers made on GWA which appeared very similiar to OP's and turned out to be clean.




so the myc didnt colonize the top of the agar, but rather dived into the agar, grew underneath the surface and then surfaced to the top again further away on the plate?

never seen anything like it and having problems picturing how or why that would happen.
too much nutes just makes my myc look like a lazy chubby.


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Re: Look at my plates... what do you see? [Re: spacechildo]
    #23393402 - 06/29/16 08:27 AM (7 years, 6 months ago)

Quote:

spacechildo said:
Quote:

Supalemonhaze said:
No lol. I'm talking about regular transfers made on GWA which appeared very similiar to OP's and turned out to be clean.




so the myc didnt colonize the top of the agar, but rather dived into the agar, grew underneath the surface and then surfaced to the top again further away on the plate?

never seen anything like it and having problems picturing how or why that would happen.
too much nutes just makes my myc look like a lazy chubby.




Not dive in per se, just colonizes the top very weakly, if you don't shine a light at an angle you couldn't see it at all. It's like it colonizes just below the surface with random linear (organized) mycelium poking through.

Then about half an inch of growth later, it surfaces hard, growing very thick and white. The following colonization is not as dense as the "surfacing" but noticeably denser than when it was growing under the surface.

To be honest, I have no idea if it was because of an overly nutritious agar recipe, it's just the only thing that varied. All my PDA and GWA batches seem to be overly-nutritious, mycelium always grows very wispy on them but I only experienced this phenomona randomly during about a 2month time span and only with the same GWA recipe.

I have been diluting my GWA with a 1:3 ratio lately, mycelium still grows wispy AF but at least it grows above the surface. I use MEA mostly, it's my favourite for thick organized growth. Never had the same results with PDA or GWA.


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Re: Look at my plates... what do you see? [Re: Supalemonhaze]
    #23393408 - 06/29/16 08:29 AM (7 years, 6 months ago)

i need to get some MEA, so i can start varying recipes for the myc. plus, i think i have been making my PDA wrong. either too nutritious or too stiff or something. im not sure if this is the case, but i just have a feeling ive been doing something wrong, for some reason hah..


--------------------
-The wise man never stops seeking knowledge.


-I wanna feel the change consume me, feel the outside turning in. I wanna feel the metamorphosis and cleansing I've endured within.


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Re: Look at my plates... what do you see? [Re: mupetmower]
    #23393413 - 06/29/16 08:32 AM (7 years, 6 months ago)

PDA is my least favourite recipe. Apart from growing only very thin and wispy mycelium, the mix is clouded by the potato flakes floating around. GWA is very clear but growth is typically the same, very hard to tell if it's organized or not and impossible to tell sectors apart. If it wasn't for MEA, I would be having a much harder time with agar.


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Re: Look at my plates... what do you see? [Re: Supalemonhaze]
    #23393434 - 06/29/16 08:46 AM (7 years, 6 months ago)

Hey...  it's all above the survace I tell you. Well, I'll transfer it anyways and keep you guys updated.
Seems to be of interest, lol.

I still don't think it's mold... except for picture 3, that could all be mold. I used PDY-agar. These potato-flakes don't dissolve fully, so maybe the nutrients in the plates aren't very evenly distributed...

SW


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Edited by Spiritwithin (06/29/16 08:48 AM)


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Re: Look at my plates... what do you see? [Re: mupetmower]
    #23393447 - 06/29/16 08:52 AM (7 years, 6 months ago)

sediments dont bother me other than for visibility, I like all agar recipes basically :shrug:
if you get real whispy growth try not diluting your gwa supa. Frank added SITR or PDA to his undiluted GWA to get it as he likes it IIRC.


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Re: Look at my plates... what do you see? [Re: Spiritwithin]
    #23393451 - 06/29/16 08:53 AM (7 years, 6 months ago)

The mix will still have dissolved flakes in it, there is nutritioun in every part of the plate. I don't think plate number 3 is mold, I think it's just wispy.

Your culture is still very young, the wierd growth could just be because there is a lot of strains growing on top of each other. Still, you should refrain from using a plate like that as an inoculant until you are completely sure it's clean. We use agar to better our chances for success, it would defeat that purpose if you were to inoculate jars with suspect mycelium.


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Re: Look at my plates... what do you see? [Re: Supalemonhaze]
    #23393509 - 06/29/16 09:12 AM (7 years, 6 months ago)

Na...  I'm not planning to inoculate anything with these plates yet. I'm just curious and it's fun for me to do this. Creating some skills too... :wink:

I have some fully colonized jars from MS sitting there waiting for their horse manure...  The plates are just a side project, since I don't see the point of cloning, but that's another subject.

I have way too much time on my hands, which is good and bad.

Thank you all for your contributions!


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Trading Golden teacher prints for Pan. cyan. prints.


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InvisibleSupalemonhaze
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Re: Look at my plates... what do you see? [Re: spacechildo]
    #23393510 - 06/29/16 09:12 AM (7 years, 6 months ago)

Quote:

spacechildo said:
sediments dont bother me other than for visibility, I like all agar recipes basically :shrug:
if you get real whispy growth try not diluting your gwa supa. Frank added SITR or PDA to his undiluted GWA to get it as he likes it IIRC.




I would think that not diluting would only make it worse. I have seen a slight improvement by changing a 1:2 ratio to a 1:3 ratio (GW:water). I guess I could try doing a small pour without diluting, just to see. A lack of nutrients should give you thicker mycelium strands though, so I dunno how that would work.

This is what I typically get with GWA. You can't see it very clearly but notice how it's impossible to tell if growth is organized or not, even witha light shining through the back. The mycelium is so thin that the light is just making it super bright.


While this is typical growth that I get with a lower nute MEA mix. (1g per 100ml) You can easily see the growth on this one, even without a light through the back. With the light, the growth is thick enough for it to not be shiny.


I will try diluting it less though, everything is worth trying at this point. I only change media every month or two, even then, I only keep it on the new recipe for only 1 transfer. I am expecting trouble if I keep doing this in the future.


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OfflineHybridprX
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Re: Look at my plates... what do you see? [Re: spacechildo]
    #23393524 - 06/29/16 09:17 AM (7 years, 6 months ago)

Quote:

spacechildo said:
Quote:

mupetmower said:
like you could still take what looks like it's growing ok and transfer and clean it up, right?




nothing looks like its growing OK to me, its all got mold mixed in.
maybe thats where our confusion came from.

water is 1/3 oxygen but breathing under water is still pretty impossible since you cant single out the oxygen molecules (or cube myc in this scenario) out of the water (contams)




Im at work sweating my ass off and I just took the time out of my day to tell you to stop talking out of your ass....there is no mold on ops plates.


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Offlinespacechildo
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Re: Look at my plates... what do you see? [Re: HybridprX]
    #23393534 - 06/29/16 09:21 AM (7 years, 6 months ago)

Quote:

HybridprX said:
Quote:

spacechildo said:
Quote:

mupetmower said:
like you could still take what looks like it's growing ok and transfer and clean it up, right?




nothing looks like its growing OK to me, its all got mold mixed in.
maybe thats where our confusion came from.

water is 1/3 oxygen but breathing under water is still pretty impossible since you cant single out the oxygen molecules (or cube myc in this scenario) out of the water (contams)




Im at work sweating my ass off and I just took the time out of my day to tell you to stop talking out of your ass....there is no mold on ops plates.




says the guy that cant even see bacteria on subs or grains :rolleyes:
I get it, you're upset we hurt you're feelings yesterday, still no reason for you to give OP bad advice :wink:


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Offlinespacechildo
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Re: Look at my plates... what do you see? [Re: spacechildo]
    #23393545 - 06/29/16 09:26 AM (7 years, 6 months ago)

Red = def mold
Blue = suspect mold

+ that thick separating line clearly showing 2 competitors meeting.


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Invisibledankington
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Re: Look at my plates... what do you see? [Re: spacechildo]
    #23393569 - 06/29/16 09:33 AM (7 years, 6 months ago)

yeah those plates are all fucked. At first the third plate looked nice because it looked so even. but I saw it on my little phone. when I zoomed in...

looks like you should focus on making those first transfers quicker, OP. Next time, try and get it before the contams catch up with the mycelium. you can try transferring to lower nutes to get the myc to 'run' faster.


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InvisibleSupalemonhaze
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Re: Look at my plates... what do you see? [Re: dankington]
    #23393584 - 06/29/16 09:37 AM (7 years, 6 months ago)

Yeah, I thought it was strange that you would have picked the last one. It looks especially weak.

LOL@ Hybrid. :smbfacepalm: Not that I'm agreeing that it is mold but he definitely shouldn't inoculate with that culture if it keeps looking like that. Space's suggestion is sound. Even I, that seen similiar growth when the culture was indeed healthy wouldn't inoculate a jar with a plate that looks like that. I would investigate more, at the very least.


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OfflinecronicrFacebook
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Re: Look at my plates... what do you see? [Re: Supalemonhaze]
    #23395259 - 06/29/16 07:18 PM (7 years, 6 months ago)

you need better plates to work with, those are going to give you a hard time.


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OfflineSpiritwithin
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Re: Look at my plates... what do you see? [Re: cronicr]
    #23401154 - 07/01/16 01:21 PM (7 years, 6 months ago)

Again, thank you all for your input. I may or may not use these plates eventually...

For the heck of it I transferred these spots:



I'll show you what will happen...


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InvisibleSupalemonhaze
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Re: Look at my plates... what do you see? [Re: Spiritwithin]
    #23402810 - 07/02/16 12:31 AM (7 years, 6 months ago)

You won't learn anything if you throw away everything that seems a bit strange. Granted, you shouldn't just put everything to grains but if you keep making transfers and a culture seems to always grow wierd, they you know something is up.


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OfflineMycoGawd
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Re: Look at my plates... what do you see? [Re: Spiritwithin]
    #23402861 - 07/02/16 01:05 AM (7 years, 6 months ago)

Looks like rhizomorphic mycelium to me. Don't see any signs of contamination.


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OfflineSpiritwithin
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Re: Look at my plates... what do you see? [Re: Spiritwithin]
    #23417032 - 07/06/16 01:54 PM (7 years, 6 months ago)

Today, 6 days later, plates in the same order. Looking good I think. Sorry for the condenstation, couldn't be helped. No contams!



--------------------
Trading Golden teacher prints for Pan. cyan. prints.


Edited by Spiritwithin (07/06/16 01:56 PM)


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