

Okay, so I cloned a C. comatus (Shaggy Mane) onto MEA plates on 9/1/14. How do I know if the growth is mycelium or bacterium? its not fuzzy, so that's not a good sign. But I transferred the tissue by stuffing it into the hole I made in the agar to cool my scalpel, maybe I can't see the fuzz because its within the agar? Kinda looks like bacterial growth, though. What do you think?







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Bacteria in my eyes! Never seen shaggy mane mycelium but I would bet money its not like that



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Does bacteria grow more vigorously than mycelium? Upon closer inspection, it seems like 4 out of the 6 plates I made have this rapid, amoebic growth while the other two have slower, stringier growth



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Pics please and yes bacteria blooms pretty fast



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the pics are in the original post. i will try to get a better picture but my phone's camera is pretty poor



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Yes, there is condensation inside the dish but immediately after transfer I seal the plates with parafilm. I'm betting the contamination came from the tissue itself because its radiating from that inoculation point out. How do I better clean my specimen before bringing it into the lab? Does anyone submerge them in alcohol?



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Edited by knomadic_niki (09/04/14 05:12 PM)



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Quote:

Juiceh said:

Looks like bacteria to me. Try taking a pic without trees and stuff visible through the agar, kinda hard to see what's going on there.











Sorry, all my windows have trees out them. Here's a few better shots. One of a bad plate (contam), one of a good plate (mycelial growth), and one that's questionable....



thanks for all your replies, i'm learning







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You only have one good plate the rest is bacteria let the good one grow chances are you may see some bacteria as well but looks good so far. If bacteria shows itself its ok this is expected transfer away from contams until you get clean plates.

Good luck and get rid of them bad ones, or watch them grow well away from where you are working and don't open them. Thats what I like to do just to watch the growth.



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cool! thanks, petri_pins!



is there a way to better sterilize my specimen before I clone it? I just washed it with water and then wiped it with an alcohol-soaked paper towel. Does anybody soak it in alcohol or anything? I'm pretty sure that was the source of my contam, even though I took the sample from the center of the cap and sterilzed my scalpel between each tissue transfer.



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Edited by knomadic_niki (09/04/14 06:42 PM)



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I wouldn't soak it, but I'll douse them with alcohol to sort of flush anything off of the surface, before ripping (not cutting) them open. Then take the smallest piece from the center.



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Yea no soak think of it like a match you want to tear in half ever so easily and get the best piece of inner flesh you can find.



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okay, thanks everybody. I just did another tissue culture yesterday of an oyster growing on a nearby cottonwood. I only did four transfers so cross fingers at least one of them works. Luckily there are plenty more where it came from if they all contam. I'm having fun learning to clone mushrooms. Next on the list: spore cultivation and strain selection....sounds little intimidating...



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Nah its easy once you get to doing it a bunch. You got a flow bench or you doing a still air box? I do flow So easy and so much free space to move around.



I'm getting nasty with my grains! Just need to set up my 55 gallon steam sterilizer.

I made a big screen drain table to dry my grains all at once. I do a 50 lb ( dry grain weight ) in one run by my self now. Its so quick and easy I just did that tonight actually with pink oysters.



I started cloning a while back,and once you get the hang of the tear and a good eye for where the meat is going to be you can get one time agar no contam clones. Thats what I been doing lately but bacteria seems to always be hiding in one or two dishes.



Strains are fun to its like racing lol what sector grows fastest first which pins first in the dish/ substrate. Its really fun and has been my growing passion.

You seem to have the bug to! Glad its going around



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Haha, I've definitely got the bug, petri_pins. I built a flow hood last year which I'd been using just for grain transfers for expansions up until a week ago. After this year's Telluride Mushroom Festival and picking the brains of scientists and cultivators for a week, I ordered some dishes, agar, malt extract, etc, and got cloning. So stoked! I can't wait until I can afford a good microscope



Hey, what part of the world do you live in? Are you cultivating indoors? I had another thread going about using the CO2 my shroom room generates to increase CO2 levels in my plant room by connecting the ventilation (with air filters, of course). Any input?



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what the hell...what forum am I in?? i was just on reddit and then magically teleported here?!



oh well! so... ya got yourself a dirty culture eh? has anyone mentioned anti-bacterial agar? or "hot pour" methods???



same thing happened with my shaggy mane culture too...it didn't go soo good, but I got a tasty oyster culture out it, had to transfer the contams away from the oyster, super aggressive lol! anyway, yay for wild cultures!!!







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haha! yea, easier to start the thread once and just link it. I should consider anti-bac agars. the already mixed (heat-stable) stuff is expensive but maybe I could buy some and add it when the agar cools.



I might have one culture that's okay, we shall see with time...



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$7.95???



and add it before your pressurecook/sterilize/autoclave /etc.



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