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Offlinetrubblesome
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Registered: 11/09/19
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Re: Let's All Grow Mushrooms 2020 [Re: Haywire] * 4
    #26388238 - 12/18/19 07:24 AM (4 years, 1 month ago)

Trubblesome's LAGM 2020
Species: Cubensis
Var: PESA

:howyoudoing:


1/1/20

Here we go.



I only did 2 plates (PESA print) because I'm a daredevil. but I also have a scope so I can verify the streak "worked." It did. Under the scope I found several locations where individual spores were just hanging out, so since I have a ton of relatively straight forward stuff going right now including a PESA plate about to go to grain and don't really NEED these to do anything in particular, I'm thinking about trying to do something fun with those and I'll be checking in on them daily. I'll use this grow as an opportunity to improve my microscope game as well.




this lone floater down below that string of other spores is the most promising one I'm keeping an eye on, but there are some others too that I might be able to slice out and pop on another dish to see what happens. probably a foolhardy endeavor but hey. let's all grow mushrooms.

1/2/2020

and we have growth!



bacterial growth. PESA 2 grew this little spot of bacteria fast, like 28 hours or so? yikes.



but this is fun, this is one of my goals for 2020 - get better at microscope work. time to bust out the stains...


1/3 early AM microscopy edition

the stain kit I got doesn't have crystal violet...I should have looked closer when I ordered but...come on. oh well. I'll wait to look at the bacterial growth, don't really have time to prep slides and shit tonight.

we do, in fact have liftoff though




none of the "loner" spores or any in the smaller groupings are doing anything yet. I'll check back on this tomorrow though, a leading edge from this could make for some fast shroomies.


1/3

I cut a diamond around that clump of spores, then sliced that diamond in half and put each half on it's own dish.










1/4: single spore transfer

I found this guy on the PESA 2 plate that's contaminated.



there wasn't another spore for a centimeter in either direction so I was confident I could pluck it and I did. This plate doesn't have too many spores on it anyway and that bacteria spot is growing rapidly so this will be my big experiment for the grow, I've wanted to do a PESA B+ cross but without obvious phenotypic differences it would be hard to verify what's happening UNLESS I can do what I've done here and isolate a single PESA spore (I'll see what F1 looks like and figure out how to deal with F2 later. problems for future me. if anyone has a thought here I'm all ears.). I'll print the biggest specimen from a B+ tub that was spawned 12/28 and do the workman cross method, scraping fresh spores in to a nearly completely colonized jar of grain from this monokaryote spore. Since a monokaryote can't produce fruits, the new spores from the print should link up with this mono mycelium, share DNA, and make fruits that have the DNA from both varieties!

SO! How I did it:

I found the spore by zeroing in on and scanning over my streak at 40x, (the max I can look at things on petri dishes under the scope, with different or thicker poured petris I can sometimes get 100x, but 40 is plenty for this). Then I lowered the condenser so that I could fit a sharpie underneath the plate. I used the light shining through from the condenser to draw a circle around where I was looking at the spore. Then I verified through the scope that this spore was the only one inside the circle. Then I cut a transfer out, working carefully to work just inside the sharpie line to be sure I didn't fuck it up.

so here it is, a single god damn spore transferred to a new plate:



I hope it germinates!! (I hope it's actually a spore too lol )

I actually had a contingency plan for this. I noticed directly on top of the sharpie line to the right of our star spore was another not quite as lonely spore, with a group of friends nearby. I figured I could get that one, and if I missed I'd end up with 2 anyway, then just for fun take the other 3 to a plate and have a 1, 2, and 3 spore plate.



I took my slice and pulled it up on the scope, ready to celebrate my amazing scalpel skills. NOPE. I missed it by about a millimeter. oh well. now I have this "5 spore" plate that I'll take to grains/a shoebox if there's anything good looking that comes out of it.



5 spore plate  :facepalm3:



The plates themselves.


race plate transfer growth:


:evil:

1/5/20



:howyoudoing:


1/8/20

on the 5th I made 2 jars of LC from both plates I'd transferred. I used the josex method more or less, though I used the tip of the needle like I was eating nachos and scooped up a nice large chunk of myc, just to make sure I had enough to get an LC started fast. If I had been more concerned about isolation or an even canopy or anything but speed I would not have done this. my method also differed in that rather than PC syringes with water in them to eject the myc poke, I sterilized a jar of RO water and drew that in to the syringes in my SAB, then flamed the needle again before performing the poke and squirt in to the LC.

By last night it looked like enough myc had grown in the LC to do some jars so I decided to do 2 from each LC.



Not what I'm looking for normally but it'll get the job done. There's millions of strands of myc in there.

First I placed a drop of each LC on a slide for observation under the scope, and a drop each on a plate as a test. Normally I would wait until these test plates show no bacteria to spawn to grain regardless of what I see under the microscope. I looked at it live and did not see any bacteria, just blown apart strands of mycelium. this does not mean the culture is clean for sure, as we're really only doing this so that if we do see bacteria, we can toss them and start over rather than wait for the test plates to show what we already know. Anyway it was good enough for me to go ahead and noc the jars. I kept these 4 jars around specifically for this, as they were a bit old and quite dry, I figured they could handle some "extra" LC to make up for the relative weakness of the LC. Well, when I went to pour the first one I poured WAY too much. probably like 50 ml (somehow the dog hair remained in the jar). So I poured the next one normally but on the heavy side still, 10-15 ml. For the next LC I figured why not pour one of these heavy too so they can be friends? so one got 50 ml, the other 10-15. here they are:



overpoured jars labeled "HEAVY"

I also swabbed some grocery store mushrooms and put them to agar last night as well. they'll definitely need a cleaning up if anything does germinate:



clockwise from top left: King Oyster, shimeji, maitake, white shimeji

UPDATE 1/10

both my LC test plates came up clean, so yesterday I took the remainder of the LC, decanted most of the liquid off the top, and dumped about 15 ml of nicely developed snot balls of liquid myc on to two new jars, one for each LC. then I took a sample from the remainder and observed it under the scope just to verify it was still clean. It looked good, but I gram stained it anyway to help me better see any bacteria that might be present. while doing that I got this pic of what I think is a clamp connection at 400x:




honestly I think the pours from yesterday will do better than the ones I poured too early, it's way more vigorous and is already showing signs of growth on the oats, the ones I poured too early are only just now showing growth. might have shot myself in the foot there trying to get a jump start! 

I also took a step back and realized that I'm probably taking on too much at once, and fucked around in my dishes too much to be actually tracking them waiting for a single spore to germ before contamination took over. so I swiped new dishes specifically for that project and I'll be growing nice looking transfers out like normal from the two transfers I already took. To be honest it's probably a waste of time and doing a fresh streak from whatever I harvest next will probably produce a single spore germination faster than the print I have. but I did it anyway because I was curious about trying out a weird method of spore application. it wasn't great but you know, not everything works: 



for the regular streaks I lightly drew my loop across some of the areas of my print with a finer spore load, then carefully turned the loop so that those spores were facing "up" on the loop. Then I streaked the plate with the bottom side of the loop touching the agar, so that the movement would more lightly draw spores off the top of the loop and increase my chances of finding single spore locations to cut from later. These were very sucessful, I found several locations with individual spores I'll be cutting out to move when I have time.

The less successful plates, labeled "scatter," I lightly drew the loop on the spore print in the same manner, but then I held the loop about 4" above the plate, and used the tip of a flame sterilized scalpel to pull back the loop and then let go, the idea being a scatter of spores would be propelled on to the plates and I could find numerous locations with single spores in easy to pluck locations. I don't think I let the plate sit long enough before moving my hands and putting the lid back on for any spores except the larger clumps to land on the surface of the agar, and scanning the surface of the plates for spores that could be literally anywhere ended up not being very fun and kind of straining on the eyes so I'll be focusing on the light streaking method for now, it works fine enough.


from the grocery store swabs, I have growth!!



everything but the maitake germinated, which is what I guessed would happen. I don't even think there are any maitake spores on that plate. I'm working the maitake in a clone method from Tradd Cotter where it's dunked underwater with just the base exposed to force growth out of the soft tissue at the base. no spores. I made a separate journal for these where I go in to that in more detail: Growing grocery store mushrooms

That's all for now - I think I'm at the point where I can just sit back and watch for a little while.

UPDATE 1/13. SPAWN TO COIR.

here we go




I kept the sub a bit on the dry side for the "heavy" box since there was so much LC in there it's certainly got enough moisture. spawned to coco at 1:1.5 with a .5 casing, so I labeled it 1:2. gave the casing a good mist and closed them up. my last run like this I harvested the first flush yesterday after 15 days. if all goes according to plan, I'll have some PESA by month's end.

UPDATE 1/16

Big day

4 single spore PESA plates, from a fresh print made and streaked sunday night.




I had found 5 spores with a decent amount of space between them I was going to watch. When I went to check them today for germination I found one that had:



So I got to it and made transfers of all 5. one of them I'm not 100% sure made the transfer so I won't include it here. I won't post them all here, but here is one as an example:



I fucked up the cut on the germinated one unfortunately, and had to put it face down on the transfer plate. hopefully that doesn't fuck it up. It also prevented me from getting a good picture of it under the scope.

I also did T1s of my king oyster, and T2s of my PESA destined for print making. I'll find one with a nice big cap and clone that and hopefully have a ton of nice big prints to make.

Update 1/17: dog shit jars
These jars are bad.




but especially the one on the left:



since they all came from the same LC there aint no way I'm spawning them to bulk. I might case the two on the right in the jar and put a baggy over the top just to see, but the one on the left gets the PC treatment. I'll give them the weekend and see how they look, but man they are dogshit, they are nasty.

:cuteshit:

but that's why I did 2 LCs. The ones from PESA LC 2112 are still lookin good in their shoeboxes.

Update for 1/20:

I cased the better 2 of the dog shit jars. The other one was PC'd for 2 hours and dumped in the garden. blech. looked like aspergillus, which tracks with what I usually find on a contam'd petri around here. I had some previously hydrated coco sitting in a cooler. It was a little on the dry side but I put it on top of the jar and gave it a good misting, just til it turned a shade darker brown. here they are:



It's not the end of the world if these don't work out.

The big part of the update though is a tek I've been working on that I spawned my remaining PESA 2112 jar to (the good LC that is in my shoeboxes, not one of the dog shit jars) and was able to document the prep process for.

Gift bags

I have some friends interested in mycology but they don't have the time, resources, or deep burning desire to make their own shit like those of us here at the shroomery might. So I thought about what a manageable way to share the beauty of growing your own mushrooms with friends might be, when I looked down at my take out container and thought it looked like the perfect size to make a little grow box out of. I tested it with some left over GT grain when I spawned a tub last month. So here it goes with documentation this time:

Get some take out containers. you don't have to be picky, anything with a lid to retain moisture will do. Far left container was something a friend brought me some short ribs in a few weeks back :crazy2: My dinner tonight came in one of the others:




1 myco quart jar of spawn makes four containers. Which means you put a 1/4 myco quart of spawn in each one. These are oats:



put about 1/2 a quart of the sub you like in there. this is straight coir after bod's coir tek.:



mix it up real nice:



I cased with a handful, or about a pint (1/2 quart) of coir:




I generously misted the tops in the manner of a shoebox, as well as the inside of the lid. For delivery, keep the lid tight in case shit bounces around. When your friend receives the gift, tell them to lightly crack 2 opposite corners to get a little gas exchange going. These are small as hell and I'd be scared they'll dry out quick if they go right to the next phase without someone who knows what they're looking for watching them. You could also wait until they've colonized and give them away at that point if you'd like to make it even easier for them.




Once the casing is colonized, they should take the lid off and put them in the plastic bag you delivered them in, give the inside of the bag a good mist, then tie it loosely and leave it somewhere with some ambient light that stays above 65F or so. A kitchen counter or something:



For the most part, they should leave them the fuck alone. I opened the bag and gave it another mist every few days, whenever I noticed the condensation on the bag walls starting to go away. Last Thursday, 22 days from spawn, I had these little buddies:



pins for flush 2 are coming in right now



spot on.

And of course the shoeboxes as posted above are going as planned. These are my 5th and 6th shoeboxes now and I think I'm finally getting the hang of them.

:mushroom2::mushroom2:GIVEAWAY:mushroom2::mushroom2:

I haven't changed my status from tired in some time. I'm fucking tired y'all. Life sucks right now. But growing mushrooms with all of you and reading about everyone's projects and plans has been a major bright spot. Part of the reason I went for the race is that shit is about to get very hectic for me and in a little while I'll probably have to take a few months off from all this. If I do win the prints I'd like to do a giveaway to pay it forward. Life being the energy you put out in to the universe, not what you take and all that.

I'll make a poll, LAGM growers can vote on a print from the contest bounty for me to grow. I'll do a shoebox at the end of summer, and I'll clone the best looking fruit from that and do a tub, make prints from it, and send those out to anyone who finishes a grow log by the end of 2020 and wants a print or two.

Update 1/22

got pins  :rockon:



trying something new to me this time. bags with holes on top to get that FAE/evap cycle right per the bod shoebox tek
my bags are kind of wack though so I stretched them down as best I could
my last few shoeboxes have had some kinda stunted growth/premature drying out/side pins so hopefully this and my more dialed in surface conditions improves the harvest.




Update 1/24:
I had some uh, questionable SAB sessions last week too. Did a ton of transfers last night for LAGM as well as my various other projects. Finally got caught up on some loose threads, hopefully cleaned up my SAB work after being really fucking lazy about it. Sorry about the pictures, fluctuating temps around here and not even 10 minutes with a hot coffee mug could get any of these clearer than what you see. Here's the LAGM stuff:

T3s:






When I tested my LC, culture 2111 had this perfect looking growth, completely uniform, I transferred it to a new plate to grow out for grain and...mold in the corner of the dish. fuck. so I took another transfer, we'll see how it does.



honestly this might be something I do from now on.
MS -> agar -> LC -> grains for tub -> tub, duh
LC test plate growth -> isolate further -> grains for tub ->  additional tub with less variety that is hopefully at least good, with an even pinset.

The King Oyster got transferred. I made 2 transfers on accident, I think one will contam - a piece of coir somehow got inside of it. what the fuck. I grabbed another plate and after transferring noticed a piece of coir stuck to condensation inside the lid!! no clue. small piece left on the table that got drawn up by static electricity as I moved the plate near it? I s2g I cleaned this fucking table top...



The Maitake swab plate started showing growth finally. Not knowing what Maitake mycelium looks like on agar, I was sure that it was going to turn green. But as the maitake clone grew out it looked similar, and both look like examples of maitake mycelium I've found online and what I've read about so what the hell why not! Not knowing which was better to take I made two transfers, one from strong cottony mycelium, and one from more whispy mycelium from further out, plus the one from the clone. I'll probably inoculate logs with it and bury them outside, but maybe I'll try one supplemented sawdust bag just for kicks. Trad Cotter details a method of progressively adding slits to bags that seems worth a shot.




swab plate up without that pesky lid in the way:


I expect these maitake transfers will need some extensive cleaning, but who knows, maybe that is all maitake mycelium in there.


Update 1/25
shoeboxes today:



"PESA 2112 1/9" spawned 1/13




"PESA 2112 HEAVY" also spawned 1/13

guessing I'll have torn veils monday night or tuesday morning. comparable to my first PESA shoebox from this print at ~15 days.

question posed to thread:
But I'm curious, is this just how PESA typically grows? or how shoeboxes typically grow? the first PESA shoebox I did had really skinny stems and the mushrooms themselves were quite short, they looked exactly like these only with a shitty pinset because it was my first shoebox and I let it dry out too much, so I thought the skinny stems were from that as well. I'm not sure if this is typical for PESA or if it's how shoebox grows typically look or if there's something I'm doing to them that I'm missing. They get misted whenever beads are just about gone but before myc is not dried out yet. They stay about 70 degrees, I removed the bags I posted about almost immediately after putting them on because they wouldn't stay standing up, and this morning I turned the lid slightly askew to give a little more FAE hoping to encourage some more stocky growth. If anyone has some insight to offer it'd be much appreciated.

Update 1/26. Torn Veils:

:rockon:












going to find a little better balance on misting/evaporating next time around, so much fuzzy foot going on.

Update 1/27. Harvest day.

Woke up to this:




harvested the ones that were ready. They dried out to this when I got home from work:



remember when I said the reckless nature of my speed run would show up in my results? There it is. 5.3 grams with 2/3rds to go. looking like a 15 gram haul from one quart of spawn.

:judyfacepalm:

I think I know why they were so skinny/went so fast - today I moved the other tub on their shelf to give it a mist and noticed how warm the bottom was. The ambient temp of the closet they're in is 70-72, but the surface of the bottom shelf, which sits above the heat duct run through the ceiling below, was at 78 according to my IR thermometer. Shit looks like what I grew on PF cakes over the summer at similarly high temps. Only that time I did it on purpose because I was a noob and thought they needed to be pretty much roasting at all times.

:facepalm3:

The other tub has more normal looking fruits, a more even pinset, should be done tomorrow (1 broken veil!), and I bet it will put out more weight. I'll do a final side by side when they're both done. The sub surface is confirmed by the IR thermometer to be at 72.

Cobweb won the race in one of the top-cased jars:



That's it! If anyone missed it, now that it's semi-official, I announced a giveaway in an earlier post, contingent on my getting to first torn veil:

Quote:

trubblesome said:
:mushroom2::mushroom2:GIVEAWAY:mushroom2::mushroom2:

I haven't changed my status from tired in some time. I'm fucking tired y'all. Life sucks right now. But growing mushrooms with all of you and reading about everyone's projects and plans has been a major bright spot. Part of the reason I went for the race is that shit is about to get very hectic for me and in a little while I'll probably have to take a few months off from all this. If I do win the prints I'd like to do a giveaway to pay it forward. Life being the energy you put out in to the universe, not what you take and all that.

I'll make a poll, LAGM growers can vote on a print from the contest bounty for me to grow. I'll do a shoebox at the end of summer, and I'll clone the best looking fruit from that and do a tub, make prints from it, and send those out to anyone who finishes a grow log by the end of 2020 and wants a print or two.




keep an eye out for that I guess!

:halfcocked:








Edited by trubblesome (01/27/20 04:35 PM)


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Offlinetrubblesome
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Re: Let's All Grow Mushrooms 2020 [Re: curious.psychonaut]
    #26411967 - 01/02/20 06:49 AM (4 years, 27 days ago)

did some things yesterday

Quote:

trubblesome said:
Here we go.



I only did 2 plates because I'm a daredevil. but I also have a scope so I can verify the streak "worked." It did. Under the scope I found several locations where individual spores were just hanging out, so since I have a ton of relatively straight forward stuff going right now including a PESA plate about to go to grain and don't really NEED these to do anything in particular, I'm thinking about trying to do something fun with those and I'll be checking in on them daily. I'll use this grow as an opportunity to improve my microscope game as well.




this lone floater down below that string of other spores is the most promising one I'm keeping an eye on, but there are some others too that I might be able to slice out and pop on another dish to see what happens. probably a foolhardy endeavor but hey. let's all grow mushrooms.




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Re: Let's All Grow Mushrooms 2020 [Re: Grimsweeper]
    #26413861 - 01/03/20 08:38 AM (4 years, 25 days ago)

heard there was prizes for first torn veil

been peepin mine through the scope, got some barely visible growth by eye, might make a transfer as well anyway

:halfcocked:



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Re: Let's All Grow Mushrooms 2020 [Re: Smartattack]
    #26413893 - 01/03/20 08:57 AM (4 years, 25 days ago)

agree with all, appreciate CBK for getting this going - definitely a fun chance to grow with a group. I have a couple other fun things planned for these plates but figure if there is a (extremely low stakes, light hearted) race going on why not try since I won't be sad if these get fucked up somehow, but I could totally salvage the streaks without much hassle if something does go wrong.

edit: hell yeah grim, cheers  :headbang:


--------------------


Edited by trubblesome (01/03/20 08:58 AM)


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Re: Let's All Grow Mushrooms 2020 [Re: AyePlus]
    #26414103 - 01/03/20 10:50 AM (4 years, 25 days ago)

fuck it. went home at lunch and made two plates from that spore clump. cut a diamond around it, then cut that diamond in half to make two plates. I came at it from the side so that the scalpel wouldn't be over anything I want to mess around with later. i figure one of these should work.



:mushroom2:


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Re: Let's All Grow Mushrooms 2020 [Re: MooseMomma]
    #26415746 - 01/04/20 09:35 AM (4 years, 24 days ago)

Quote:

MooseMomma said:
Even though I am definitely getting a rough start, lm determined to get the fuckers to grow. I am working with Psilocybe Cubensis Blue Meanie purchased from a vendor in a syringe, which was contaminated with what I am assuming is yeast. They were the only spores I had on Jan 1 because I am really new and I’m waiting for my first monotub to produce my very first flush (KSSS). I’m definitely glad I acquired spores from a variety of sources! The Blue Meanies are going to be a challenge. I also had some good luck with Psilocybe Cyanescens which I am growing mycelium to cultivate in an outdoor bed. I did a spore solution on Agar with them and have done very well. I have some LC of Penis Envy and KSSS that’ll inoculate grain bags and BRF jars tomorrow. I am also working with Blue Oysters, Maitake, and Chanterelles. I’ve been at this for about 6 weeks and these forums are awesome!
Does anyone know what organism this is?




I'm sorry you got some infected shit, that sucks! are you inoculating your dishes by just putting a drop of spore syringe on or are you streaking with a loop?


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Re: Let's All Grow Mushrooms 2020 [Re: Jarhead3521]
    #26416579 - 01/04/20 07:38 PM (4 years, 24 days ago)

Got some stuff done tonight, details here: some single spore transfer attempts

highlight is this lone spore I found and transferred. goal is to workman method it with a big B+ print if it germinates.



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Re: Let's All Grow Mushrooms 2020 [Re: A.k.a]
    #26416854 - 01/04/20 11:27 PM (4 years, 24 days ago)

Quote:

A.k.a said:
When you guys swipe with a loop does it do anything to the agar?

Idk if it’s cuz my plates are too hard or what but it seems like there’s either no marks at all or the agar splits all the way down to the bottom of the dish.

I poured some softer ones for the morning so I need to learn how to use a loop better.





when your loop moves across the agar (which appears smooth but at the level of the loop is quite bumpy) the spores are shed from the loop - most don't even land in the direct path where the loop touched the agar - you can see these spores (circled in red) from my streak that are about half a millimeter above where the loop streak is, circled in green:



even if you can't see it yet, they're probably germinating at a turn in the streak where there are clusters of them:



you can see the streak even more clearly here, as well as all the spores in the bend of the path. you can trust that if you at all agitated the surface of a print and then touched the agar or even moved it around near the surface, spores were deposited. You've mentioned a couple times "hard agar," what was your agar recipe?


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Re: Let's All Grow Mushrooms 2020 [Re: A.k.a]
    #26416882 - 01/05/20 12:01 AM (4 years, 24 days ago)

oh wow yeah that's frustrating! back in the day when it was my job to pour plates we mixed at 20 g agar/L so I think you'll be alright


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Re: Let's All Grow Mushrooms 2020 [Re: tealseal]
    #26417113 - 01/05/20 07:15 AM (4 years, 24 days ago)

Quote:

tealseal said:
Quote:

trubblesome said:
oh wow yeah that's frustrating! back in the day when it was my job to pour plates we mixed at 20 g agar/L so I think you'll be alright




shit, i sometimes mix 50g to a L
thats wat my jar says, i just been doing 25g to 500ml




I'm guessing you're using agar flakes then, which are typically instructed to mix at about twice the rate of agar powder


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Re: Let's All Grow Mushrooms 2020 [Re: eatyualive] * 1
    #26418579 - 01/05/20 11:26 PM (4 years, 23 days ago)



:howyoudoing:


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Re: Let's All Grow Mushrooms 2020 [Re: Greg] * 1
    #26420057 - 01/06/20 09:14 PM (4 years, 22 days ago)



I'd like to introduce my new LC tek, hair of the dog. as you can see, caramelized malt sugars cling to the stray dog hair in the jar, making for something of a life raft for strands of myc floating through the culture. allowing the dog hair to collect myc strands and settle until new growth is visible before breaking up the strand with the stir bar again allows for rapid culture growth and a guaranteed increase in potency.



JK JK
BUT FOR REAL I LOVE HIM BUT HOW IS THERE NO PLACE I CAN ESCAPE THE DOG HAIR??


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Re: Let's All Grow Mushrooms 2020 [Re: verum subsequentis]
    #26420066 - 01/06/20 09:18 PM (4 years, 22 days ago)

I've made LC at home a couple times now

I make 2 jars each time. without fail every time, one of them has a fucking dog hair in it.

if it tests aight you know I'm using it though :crazy2:


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Re: Let's All Grow Mushrooms 2020 [Re: Nobler Hino]
    #26420501 - 01/07/20 07:29 AM (4 years, 22 days ago)

I was mostly kidding but strands of organic material tangling by chance and their importance for life/the universe/existence has been kind of a theme of the last 2 trips I've taken so here's my opportunity to sound pretty out there.

I do think that there's something to it catching bits of caramelized LME as it swirls around (like smartattack said, a scaffold, kind of like phosphate and deoxyribose in DNA :laugh:), that in turn catch bits of mycelia that are floating around. as it swirls and finally settles on the bottom of the jar, those bits grow out some as they sit undisturbed, and make use of the sugars attached to the hair as a food source. when I hit the agitation bar again they all get exploded and the process begins again. this caramelized sugar stranding/clumping happens naturally, but to a lesser degree in my other jar, though many of the caramelized bits end up getting stuck to the side of the jar above the water line, so maybe the dog hair does play a role in keeping them floating in the solution. I'm definitely not concerned about its safety since I know it was in the jar before the PC, and even though I'm typing right now that I'm not going to put dog hairs in an LC on purpose, as I type it I think "well, maybe that'd make a good thread though..."


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Re: Let's All Grow Mushrooms 2020 [Re: MooseMomma] * 1
    #26421218 - 01/07/20 02:22 PM (4 years, 21 days ago)

I'd also suggest acquiring or making your own loop. put a squirt of that spore syringe in a clean shot glass or an unused sterile petri dish, then dip a (sterilized by flame and then cooled) loop in it and draw an S/Z pattern with it across the plate so that the spores and the bacteria can get more separation.

re: soot, ime soot is the product of incomplete combustion from whatever fuel your heat source uses. soot = needs more air. a properly dialed in burner/torch/whatever won't leave soot. A fisher burner is the tool I miss the most from the lab, but I'm not quite at the point where I'm gonna pipe gas to my mush cult space.


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Re: Let's All Grow Mushrooms 2020 [Re: eatyualive] * 1
    #26422532 - 01/08/20 08:40 AM (4 years, 20 days ago)

updated my first post

the gist is that I put a lot of LC on to grain last night. accidentally sloshed 50 ml in to one quart jar, so I did another like that as well for consistency's sake. other 2 jars were on the heavy side too but nothing wild. 10-15 ml. the grains were very dry specifically so I could inoculate heavy but not 50 ml heavy!




swabbed grocery store mushrooms to agar last night as well:



King Oyster, brown and white Shimeji, Maitake


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Re: Let's All Grow Mushrooms 2020 [Re: Inthepit] * 1
    #26422729 - 01/08/20 10:50 AM (4 years, 20 days ago)

Quote:

Inthepit said:
I hope this isn't too far off topic, just wanna ask a quick question.
Quote:

trubblesome said:
swabbed grocery store mushrooms to agar last night as well:



There's some great shrooms in the market here in France. Do you swab the gills of the mushroom or try for a print from it?
A lot of them haven't even opened the veil.





this was my first time ever doing it so I can't say this is the best or even a great way to do it, but I tried to print the shimeji and oyster - only the oyster dropped anything, so i swabbed the gills of the gilled ones (kind of aggressively, figuring they'd been harvested who knows how long ago), the pores of the maitake, and aggressively dragged/rolled the swab around on the plates. I also cut clones from interior tissue just above the gills, and from near the base of the maitake just to see how that might work too.

the only one I'm confident I'll ever see fruits from is the oyster, but this seemed like a fun thing to try, LAGM really has me in the spirit!

I also expect to grow a lot of other stuff on these plates and have a lot of clean up to do considering the journey the fruits made to my fridge


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Re: Let's All Grow Mushrooms 2020 [Re: ComebackKid] * 1
    #26427137 - 01/10/20 07:12 PM (4 years, 18 days ago)

Update 1/10:

both my LC test plates came up clean, so yesterday I took the remainder of the LC, decanted most of the liquid off the top, and dumped about 15 ml of nicely developed snot balls of liquid myc on to two new jars, one for each LC. then I took a sample from the remainder and observed it under the scope just to verify it was still clean. It looked good, but I gram stained it anyway to help me better see any bacteria that might be present. while doing that I got this pic of what I think is a clamp connection at 400x:




honestly I think the pours from yesterday will do better than the ones I poured too early, it's way more vigorous and is already showing signs of growth on the oats, the ones I poured too early are only just now showing growth. might have shot myself in the foot there trying to get a jump start! 

I also took a step back and realized that I'm probably taking on too much at once, and fucked around in my dishes too much to be actually tracking them waiting for a single spore to germ before contamination took over. so I swiped new dishes specifically for that project and I'll be growing nice looking transfers out like normal from the two transfers I already took. To be honest it's probably a waste of time and doing a fresh streak from whatever I harvest next will probably produce a single spore germination faster than the print I have. but I did it anyway because I was curious about trying out a weird method of spore application. it wasn't great but you know, not everything works: 



for the regular streaks I lightly drew my loop across some of the areas of my print with a finer spore load, then carefully turned the loop so that those spores were facing "up" on the loop. Then I streaked the plate with the bottom side of the loop touching the agar, so that the movement would more lightly draw spores off the top of the loop and increase my chances of finding single spore locations to cut from later. These were very sucessful, I found several locations with individual spores I'll be cutting out to move when I have time.

The less successful plates, labeled "scatter," I lightly drew the loop on the spore print in the same manner, but then I held the loop about 4" above the plate, and used the tip of a flame sterilized scalpel to pull back the loop and then let go, the idea being a scatter of spores would be propelled on to the plates and I could find numerous locations with single spores in easy to pluck locations. I don't think I let the plate sit long enough before moving my hands and putting the lid back on for any spores except the larger clumps to land on the surface of the agar, and scanning the surface of the plates for spores that could be literally anywhere ended up not being very fun and kind of straining on the eyes so I'll be focusing on the light streaking method for now, it works fine enough.


from the grocery store swabs, I have growth!!



everything but the maitake germinated, which is what I guessed would happen. I don't even think there are any maitake spores on that plate. I'm working the maitake in a clone method from Tradd Cotter where it's dunked underwater with just the base exposed to force growth out of the soft tissue at the base. no spores. I made a separate journal for these where I go in to that in more detail: Growing grocery store mushrooms

That's all for now - I think I'm at the point where I can just sit back and watch for a little while.




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Re: Let's All Grow Mushrooms 2020 [Re: trubblesome]
    #26430196 - 01/12/20 07:11 PM (4 years, 16 days ago)

Quote:

trubblesome said:



from the grocery store swabs, I have growth!!








update 1/12:

this was all mold except the oyster. "lol"



so for now the King Oyster will be the only gourmet-from-spores I'll be doing from that batch. I just picked up another mixed pack though and it's got chestnut/velvet pioppini/whatever you want to call it in it so, why not try and grow those too? why the fuck not. throw it in there. maybe tomorrow.

tonight I'm prepping coco for PESA shoeboxes over the next few days, starting with the HEAVY jars which should be ready to go come AM. I love LC because it just goes and goes looking like normal ass grains, then BOOM! myc all over.


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Re: Let's All Grow Mushrooms 2020 [Re: Jooby]
    #26431860 - 01/13/20 05:49 PM (4 years, 15 days ago)

here we go




I kept the sub a bit on the dry side for the "heavy" box since there was so much LC in there it's certainly got enough moisture. spawned to coco at 1:1.5 with a .5 casing, so I labeled it 1:2. gave the casing a good mist and closed them up. my last run like this I harvested the first flush yesterday after 15 days. if all goes according to plan, I'll have some PESA by month's end.


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