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OfflineGrenik
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Re: Check out my agar, please? [Re: karri0n]
    #27040920 - 11/15/20 06:53 PM (11 days, 6 hours ago)

Nice to see you getting a work process that works for you. Just to set some expectations...not every drop will have spores in it. I normally do 4 plates from a syringe and 1 or 2 normally have no growth.  No contamination, no mycelium, just a blank plate. But on the plates that do grow, I always get a good area I can transfer from.

Good luck to you!


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Offlineblack strat
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Re: Check out my agar, please? [Re: Grenik]
    #27041053 - 11/15/20 08:23 PM (11 days, 4 hours ago)

Quote:

Grenik said:
Nice to see you getting a work process that works for you. Just to set some expectations...not every drop will have spores in it. I normally do 4 plates from a syringe and 1 or 2 normally have no growth.  No contamination, no mycelium, just a blank plate. But on the plates that do grow, I always get a good area I can transfer from.

Good luck to you!




yes! this is what really had me thinking last night. I couldn't imagine EVERY drop having spores, so doing needle -> loop -> plate just didn't seem totally guaranteed in my mind. glad you confirmed and gave realistic expectations.

I know I said a few days for a pic lol but I wanna post it now.



just based on my 'luck' so far, this isn't mycelium, is it? :sad:

karri0n, took your advice - with the new plates I'm leaving them where they are now, undisturbed. but I do see the zigzags are already little bacteria-boulevards.. I was a lot more careful this time around too, so I dunno men....

I DO have a couple of prints coming in the mail now though. I wanna try everything lol. this shit is so cool.

thanks again bros.

oh, super amateur question - for an iso spray bottle -- do y'all dilute with water? any ballpark-ratio you could give me please?

and next session i am DEFINITELY gonna try the sterile swab way. just bought 300 cheapy qtips, gonna wrap in foil n PC.... is there a tek for it, or do ya jus sorta make little foil packages n elevate them in PCer, and bada-bing, sterile swabs?

:mushroom2:


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OfflineGrenik
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Re: Check out my agar, please? [Re: black strat]
    #27041064 - 11/15/20 08:33 PM (11 days, 4 hours ago)

I think there is a good chance that you have something salvageable. I would make a transfer where I have indicated and try growing it out to see what happens.



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Offlineblack strat
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Re: Check out my agar, please? [Re: Grenik]
    #27041120 - 11/15/20 09:11 PM (11 days, 3 hours ago)

really?? ok, will do, thanks brother.

damn, so that's not mold, huh? it's quite fuzzy.  then again, I don't know jack shit. hah :thumbup:


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Offlinekarri0n
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Re: Check out my agar, please? [Re: black strat]
    #27041211 - 11/15/20 10:30 PM (11 days, 2 hours ago)

Quote:

black strat said:
is there a tek for it, or do ya jus sorta make little foil packages n elevate them in PCer, and bada-bing, sterile swabs?

:mushroom2:




I didn't see a tek when I did it, just a description. I stuck em in a jar with 4 micropore-covered GE holes, just because I had it sitting around from pf tek. Then PC'd for 20 mins. Then I did see a tek today which basically mirrored the process I used:

https://www.shroomery.org/forums/showflat.php/Number/22556356/fpart/all/vc/1


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Edited by karri0n (11/15/20 10:31 PM)


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Offlineblack strat
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Re: Check out my agar, please? [Re: karri0n]
    #27042782 - 11/16/20 09:30 PM (10 days, 3 hours ago)

good evening fellas.

jus PCed 400ml of new agar, waiting for it to come down n hit that sweet temp to pour, fuck, I think Bods tek says 110F? whatever, gotta go look.  wrapped some qtips too.

anyway, please forgive my obsessive questioning, but are we sure this is myc? and not some devious fuckin fuzzy white mold?  and if we're set to go, do you concur with my red marks on where to slice from? not so much on the smaller plate, I think that one should grow a little more.. but at least from the bigger, more occupied plate?

(6 pics, but we're looking at jus 2 separate plates here)



thanks.


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OfflineGrenik
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Re: Check out my agar, please? [Re: black strat]
    #27043007 - 11/16/20 11:28 PM (10 days, 1 hour ago)

Are all of your plates from the same source?

I am still a newbie, I just swabbed my 550th plate tonight. I feel confident that you are in good shape and it is worth transferring. I thought from the beginning you just needed time, but I am still learning also.

I find that mycelium grows with that leading edge that you have in your pics. I find contams to have a smoother, more uniform leading edge. I am sure it is not a perfect way to identify, but that growth looks good to me.  I will try and post some magnified shots so you can see the difference, but it may be a few days before I get to it. Also, mycelium seems lighter and cotton like while contams are more matted and dense.

Still, I am always happy when mushrooms grow or my plates become rhizomorphic. That is the only way I really know.:confused:

I like your choice of transfer choices. I would try the ? location for practice. On the plate with 4 red marks, I would also try the 6:00 position. You could drop the 3:00 one if you only want to do 4. But your original choices are also good spots, so your choice.


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Offlinescarabaeus
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Re: Check out my agar, please? [Re: black strat]
    #27043029 - 11/16/20 11:37 PM (10 days, 1 hour ago)

It looks to me like you have the right idea, i.e. always transfer AWAY from the contamination. It is obvious to you by now (and everyone else  lol), that you have a bacterial mess. I have salvaged plates just as bad by transfer though, so I say give it a shot.

I always have a jar of anti-bacterial agar on the shelf just for this kind of situation. It consists of the usual agar recipe with the addition of gentamycin sulfate (g sulfate is the anti- bacterial component). It is available as an agar supplement from one of the Shroomery vendors (I can't remember which one at the moment). It is a bit of a pain to mix if you are not familiar with this kind of thing but definitely doable. Premixed anti-bact. agar is available from Paul Stamets' company but I don't do business with him because I think his business practices suck -but that is a topic for another thread.

The above paragraph was not targeted specifically for you, but rather just as some general info for whoever may read this. You may still be able to salvage and be  a winner using what you currently have to work with. Good luck! :mushroom2:


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Offlinescarabaeus
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Re: Check out my agar, please? [Re: scarabaeus]
    #27043041 - 11/16/20 11:44 PM (10 days, 1 hour ago)

Oh yeah! I forgot to mention. If in the future you have a mold contamination rather than a bacterial one, in my experience it is better to just start over. A mold that is turning colors (usually green) means that it is making spores, and spores float around and get everywhere. At least bacteria doesn't make a 'contamination fountain' like a sporulating mold will.


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OfflineGrenik
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Re: Check out my agar, please? [Re: scarabaeus]
    #27043044 - 11/16/20 11:45 PM (10 days, 1 hour ago)

You can get it here. You can also buy plates from several sources with it already added to the agar.

How did plates 8 and 12 do from the original post?


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Offlinescarabaeus
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Re: Check out my agar, please? [Re: Grenik]
    #27043057 - 11/16/20 11:56 PM (10 days, 1 hour ago)

Ah yes, shroom supply. I was kinda thinking it was them but wasn't sure. BTW, I have ordered stuff from them in the past and have found them to be quick and reliable. Thanks Grenik :thumbup:


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Offlineblack strat
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Re: Check out my agar, please? [Re: scarabaeus]
    #27043097 - 11/17/20 12:43 AM (10 days, 26 minutes ago)

you guys are the shit. I'm really digging this community so far. there is SO much information on this site, it's kinda fuckin absurd.

Grenik, yes sir, the substance in question is in fact 'light, and cotton-like' by the looks of it! almost wispy hah. also, when you say 'from the same source', what do ya mean exactly? I poured all 13 of those together, yes, and inoculated the same time, but I used 4 different syringes. I don't know why, I wanted to try em all and see what would happen. there's GT, Dancing Tiger, Brazil, and Mexican Dutch King. the 2 I'm transferring from are both Dancing Tiger. I poured 30 fresh cups of agar tonight, so tomorrow I'll transfer these. thanks for the advice brother.

scar, I had no idea that stuff existed, I will definitely be ordering some. and yeah, it is a mess lol, but it's my first time and fuck it, maybe what better way to start off than trying to see if I can climb my way out of this mess, ya know? to be completely honest, I actually am having a good time with this shit. yeah, of course it'd be awesome if this go-around actually pans out for me, but it's been fun so far either way. and thank you for your input as well man!!

:mushroom2: :mushroom2: :mushroom2: :mushroom2:


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Offlineblack strat
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Re: Check out my agar, please? [Re: Grenik]
    #27043100 - 11/17/20 12:47 AM (10 days, 21 minutes ago)

Quote:

Grenik said:
You can get it here. You can also buy plates from several sources with it already added to the agar.

How did plates 8 and 12 do from the original post?




hmm, honestly not too sure which is which anymore exactly :/ most of them are ONLY snot still. the 2 above I guess are showing promise, and I'd say there are another maybe 2 or 3 that I'm holding out jus a little bit of hope for, and they're jus running a little late to the party.


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InvisibleIdiotCircusBoy
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Re: Check out my agar, please? [Re: black strat]
    #27043367 - 11/17/20 08:56 AM (9 days, 16 hours ago)

If that is mycelium you need to transfer asap.


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Offlinekarri0n
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Re: Check out my agar, please? [Re: IdiotCircusBoy]
    #27044150 - 11/17/20 06:17 PM (9 days, 6 hours ago)

I really hope that's not mold.

I have some growth that looks just like that on one of my plates and I Identified it as mold because it grew straight from the edge and quickly, and it has that hairy "brillo" look.

I do my photos up through the bottom of the plate since my no-pours don't have great clarity and are tough to see on camera through the tops and sides, but I will explain what I am talking about best I can.



This grew on this plate 1 day after inoculating my plates, starting from the far left edge in the first pic and from the bottom in the second. The two little circles or "bubbles" are new, and might be bacteria, or even mushroom mycelium that germinated under the mold colony.



Side view:


Note how brillo, wispy, hairy it looks, and also how quickly it grew. It appeared as a tiny spot in the corner, two days before the mycelium in the following pics appeared multiple places, and has consumed almost the whole plate, while the mycelium has the one larger colony but is mostly moving slower.



Side view:


The mycelium is also cottony and fluffy. However it seems to be thicker, and doesn't have a brillo hairy appearance moving in wild directions. I also notice that cubensis mycelium tends to eat the food coloring out of the agar, turning it to its natural brown color(I used grainwater, it will be white if you used flour) and mold doesn't.

I really hope I was able to detail the difference in appearance here. I don't know that what you have is mold or mycelium, but I wish you the best of luck and just hope I can help.

The biggest way to tell for me is speed of growth, because cubensis mycelium appearance really can be pretty variable. Mycelium is slower than mold.


Edited by karri0n (11/17/20 06:32 PM)


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Offlineblack strat
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Re: Check out my agar, please? [Re: karri0n]
    #27044441 - 11/17/20 09:17 PM (9 days, 3 hours ago)

fuck....

man, I just really really do not know. I did the transfers a few hours ago.  all I can say for a fact is that at the very outermost edges of my 'white stuff', it begins to straighten out. I have no clue if that means anything. also, it always seems to be growing away from the bacteria. damnit, who the fuck knows. I don't even know how I am supposed to tell.


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Offlineblack strat
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Re: Check out my agar, please? [Re: black strat]
    #27044454 - 11/17/20 09:26 PM (9 days, 3 hours ago)

yeah, now that I'm looking online at pics, I think it's cobweb mold. awesome.

basically just wasted 15 cups of agar carefully transferring different pieces of cobweb mold. wow. I am saaaalty.

edit: ok, so here is one of my newer cups, I swiped with an inoc loop a few days ago..


if what I had earlier is cobweb, then this most certainly is too. it's growing/looking EXACTLY how those others did at first when they started. I don't fuckin get it. how can none.. out of like almost 40 cups.. none of them give me even a hint of myc??

fuck. I'm done for the night. this is such a bummer.

edit again: jus read that putting peroxide on cobweb mold eats at it like wet cotton candy. dunno how accurate that it. swabbed some peroxide on the original cups I took the transfers from.... nothing happened. jus sat there kinda. who knows. who the fuck knows lol.


Edited by black strat (11/17/20 10:25 PM)


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OfflineGrenik
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Re: Check out my agar, please? [Re: black strat]
    #27044562 - 11/17/20 10:43 PM (9 days, 2 hours ago)

Hmmm

You are probably putting too much material on the plate. One drop on the inoculation loop and spread it around. A typical inoculation loop holds 10 microliter of liquid.  So 1 mL would be enough for 100 plates. 1 drop is about 50 microliter, so 5x the amount you need. 

Mycelium grows and spreads. It may start slower than mold, but it normally spreads faster than contams. You want a very small sample and let the mycelium run, then transfer from the edges.

Let’s look at those transferred samples in a week. The original plates that you swabbed with peroxide will probably die, that does not prove cobweb, it proves peroxide is bad. 

Deep breath.

Switch to spore prints if the syringes concern you.


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Edited by Grenik (11/17/20 10:46 PM)


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Offlineblack strat
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Re: Check out my agar, please? [Re: Grenik]
    #27044637 - 11/17/20 11:50 PM (9 days, 1 hour ago)

:tongue2:

I was dying reading that reddit post. especially because I AM that noob hahah. man, I feel dumb. but you know, the only reason I went with the ol cobweb theory is because shroomscout's right - it keeps getting repeated over and over, propagating that paranoia, then my stupid ass sees all that, and suddenly "wahh, I have cobweb!!" lol, oh boy.

alright. I'm good. gonna leave em alone like you said. and yeah man, my prints should be here any day now, can't wait. thanks bro.


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Offlinekarri0n
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Re: Check out my agar, please? [Re: black strat]
    #27044679 - 11/18/20 12:53 AM (9 days, 16 minutes ago)

Stay away from peroxide. It's toxic to any fungus, including mycelium.

Those swipe Plates still look like too much water to me.

Keep an eye on your swabbed plates and your transfer plates. Have patience.

As long as you are putting very, very small amounts of spore solution on the plate, and following sterile technique, you will get mycelium.

I saw you say something about being worried that not every drop will have spores. A drop will have thousands of spores. If you see them in the water, those are actually clumps. They're microscopic. To give you a hint of how many are in there, I aspirated sterile water back into the spore syringe after it was empty to rinse the last leavings out, put that on a BRF cake, and it germinated. This was just testing to see if there really were still spores in there.

Are these plates different varieties of cubes? You should see slightly different looking mycelium between varieties and between plates.

At least a few are going to have usable growth. You're gaining experience and understanding with every step, so just have patience and take heart in the fact that you're both learning and will see success.


--------------------
Anything written above is just as likely to be accurate as it is to be corrected by someone smarter than me two posts from now.

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