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InvisibleMicroMycelium
Curious Optimizer

Registered: 06/06/23
Posts: 461
Loc: Probably at work
First Grow- Tidal Wave MultiSpore * 5
    #28350605 - 06/07/23 08:38 AM (7 months, 17 days ago)

A log of my first experience growing cubes.
Purpose of this is to record data throughout the process and throw out my thoughts & questions if any vets want to chime in.

As I have harvested my first ever fruits, I would like to thank everyone who chimed in here and in other threads as well. This site and the people on it have been a tremendously valuable resource.

I'm mild OCD and major ADHD so excuse any over-engineering. I intend to learn along the way and will likely make mistakes as lessons for the future.

1. Everything is housed in my grow tent for privacy and containment.
2. 7500k Grow lights on a 12/12 cycle are zip tied to the racks above the tubs.
3. Fruits for the log will be born from 2 MSS I got from an online vendor.

My Initial Set Up:


My Most Recent Setup:
I moved away from the Max Yield Bins and stick with 6qt shoeboxes to dial in genetics before larger tubs. Moving to 12qt shoeboxes in my next phase.



Shoebox Set up:
After pinsets have formed and begin to reach the sterilite tub lids I’m placing upside down boxes on top to allow light penetration to direct growth and additional room for fruits to grow.


Agar Notes:
Originally I started with premade PDA Agar Plates which I mistakenly did not streak and only allowed the solution to roll around on.





I stuck with PDA plates for initial transfers.



Growth on these was fairly decent and no visible satellite contamination formed through their colonization however the mycelium growth was not very uniform.



Aside from a few plates which came out awesome.


I got creative and decided to make a high-ER nutrient Agar to compare to my PDA plates.
MEPYAC Agar: This is Malt/Peptone/Yeast/Charcoal Agar.
Recipe:
- 500ml distilled water
- 9g Agar
- 7g Light Malt Extract
- 0.5g Peptone
- 0.5g Nutritional Yeast
- 1g Activated Charcoal




This recipe tremendously slowed the mycelium growth and caused many inconsistencies even after multiple transfers and cutting the total nutrients down by 40%~ in a second batch.





I decided to add in HG PP5 plates due to their lessened need of parafilm.
Here were my best plates from this crappy formula.



Ultimately I have most recently gone back to a simpler recipe.
MYA Agar:

-10g agar/7g Light Malt/0.5g Nutritional Yeast.
Aside from the sediment the nut. Yeast leaves, it appears to be more conducive to germination.




Grain Prep:
I have chosen WBS as my first choice of grains.
My first attempt with prepping the WBS was the flash-boil prep method.
-Rinsing the wbs a few times until water is clear
-Boil a pot of water, turn off the heat
-Air dry grains until they don’t stick together
-Place WBS in water with a lid and let set for 30-45 minutes.
-Jar the WBS and PC at 15 psi for 2.5 hours.
I inoculated the jars via agar plate slices with a flame-sterilized scalpel in my SAB.






I noticed the grains to be too dry with a few stalled out jars using this method.
Here is an example of a colonized-yet stalled jar from being too dry:

In attempt to “reinvigorate” the jar I put some tap water in a mason jar and ran it through the PC’d at 15psi a few times and poured roughly 20ml of sterile water in the jar.
I then shook the jar once every 24hr for 3 days resulting in the mycelium density exponentially increasing.
Looks slightly bacterial, but fruiting opportunity is greatly increased with stronger mycelium obviously so I spawned in anyways.


A few of my other jars for reference:







After experimenting my future WBS preps will include a 24hr soak and an optional 10-20 minute simmer if still not hydrated to my liking.
Due to the grain inconsistencies I will be switching to whole race oats for my next grain jars.

Initial Shoeboxes Progression:
Utilizing my 6qt shoeboxes I mixed 1qt colonized WBS with 2qt CVG, slightly compacted, a loose pseudo-casing of roughly 0.5qt CVG, misted the casing, and left the lids on until pinsets formed.










Added a few more tubs.


First pins have begun to form 16 days into spawning



Pins begin to take shape 18 days in.


19 days in and this fruit is growing rapidly. I pulled it for a clone due to this however many people argued it was an abort. The tubs I will spawn in the future with its genetics will tell.



21 days into spawning and the difference between the two tubs is very apparent.




23 days in and the fastest growing fruits have begun to crack.
Some said this was due to to much FAE.


Upon comparing the flushes’ fruits in their entirety, I determined that was not the case after sealing the tub’s creases and comparing it the simultaneously developing tub. I attribute the cracks in this batch due to their rapid growth as the fruits in this shoebox grew twice as large and twice as fast compared to the other.
Compared run at the same age and conditions:


25 days in.


26 days into spawning, my first multispore flush.


I then bottom-watered about half way with distilled water 2 days post-harvest(28 days in) due to lack of additional pins forming.

Now 7 days after soaking the substrate, 35 days in, second flush has developed.



36 days in and my second flush is ready and harvested.



While there are still a few pins slowly developing, I have harvest two flushes from each 6qt shoebox. Total wet yield was roughly 550.

Additions Not Yet Being Utilized:
- 2x 20qt Max Yield Mono Tubs
- 2x 120ml CPU fans
- 2 Cyclical timers for the fans for FAE
- 1 reptile fogger
- Ink Bird Humidity Sensor

Future clones:
Due to my first cube grow being a multispore lottery, I took tissue samples from the fruits I favored for most consistent future grows.
My process for these clones was as follows:
-Harvest Fruit based on characteristics
-Manually split fruit inside SAB.
-Use flame sterilized scalpel to take sterile tissue sample from inside of stipe/stem to avoid spores and contam
-Placed tissue sample on MYA agar in SAB.
-As agar germinates, I have water agar HG plates on standby to clean them up if necessary.

Here are my cloned fruits:

Clone 1: very first pin to form, formed and grew faster than any other fruit.


Clone 2: Fruit with the thickest stem, TW genetics present with wavy cap.


Clone 3: Most overall decent fruit. Tall & thick.



Clone 4: Twice as tall as any other fruit, leucistic variety with lighter colored cap.


Clone 5: conjoined based(cluster?), hearty, umbo point on on cap.


Edited by MicroMycelium (08/14/23 12:35 PM)


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InvisibleWay
The


Registered: 01/14/23
Posts: 4,336
Loc: A long way away
Re: First Cultivation Log with Tidal Wave MSS [Re: MicroMycelium] * 2
    #28350672 - 06/07/23 10:26 AM (7 months, 17 days ago)

I hope to see you succeed. It's pretty difficult to grow cubes well in a tent from my understanding.

Good job starting with agar :thumbup:

Welcome by the way.


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

That's the way she goes, boys. Sometimes she goes, sometimes she doesn't, cause that's the fuckin way she goes.


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InvisibleMicroMycelium
Curious Optimizer

Registered: 06/06/23
Posts: 461
Loc: Probably at work
First Agar Transfer [Re: MicroMycelium] * 1
    #28351072 - 06/07/23 03:49 PM (7 months, 17 days ago)

Since my MSS made an unorganized mess of mycelium all over the agar plates I did my first ever Agar-to-Agar transfer today. This was done 9 days after introducing the spores.

Here is the bottom of both my initial plates 9 days in, both from different Tidal Wave MSS.
-Syringe 1:

-Syringe 2:



The transferred sections:




And for fun with the lid off to examine the development...
Looks mostly fluffy(Tomentose?) but I tried to take the most root-looking(rhizomorphic?) sections.
Also I don't see any contamination? :thumbup:



Edited by MicroMycelium (06/07/23 03:51 PM)


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InvisibleMicroMycelium
Curious Optimizer

Registered: 06/06/23
Posts: 461
Loc: Probably at work
Did a few more transfers [Re: MicroMycelium] * 1
    #28353932 - 06/10/23 12:02 AM (7 months, 15 days ago)

I pulled a few more transfers from my original spore plates. In attempt to get more genetic samples to choose from when I inoculate my grain bags and LC.

You can see the original transfers 2 days after being selected are starting to colonize their plates without contamination.






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InvisibleMicroMycelium
Curious Optimizer

Registered: 06/06/23
Posts: 461
Loc: Probably at work
Little additions [Re: MicroMycelium] * 1
    #28362559 - 06/16/23 08:26 PM (7 months, 8 days ago)

Since I’ll be transferring my agar a few times, trying to get the best culture I can, I felt it would be wasteful to leave the plates I took my sectors from.
So I decided I will be doing a few 6qt shoebox teks with the old plates to “practice” before I colonize my big monotubs.

Today I made some WBS grain spawn that’s currently in my PC as well as  my first attempt at mixing my own agar(MEPYAC).
Will post some photos and agar formula once it all cools from my PC and I can handle it :thumbup:


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InvisibleMicroMycelium
Curious Optimizer

Registered: 06/06/23
Posts: 461
Loc: Probably at work
WBS Grain Spawn [Re: MicroMycelium] * 1
    #28362763 - 06/16/23 11:10 PM (7 months, 8 days ago)

WBS grain spawn

-1qt Mason jars with no-leak lids
-Roughly 850g WBS each

Process

1. Put all the grains in a pot and sifted/rinsed with cold water 7-10 times until the water was relatively clear.
2. Pulled 99% of the sunflower seeds floating on the water out.
3. Brought a pot of water to a boil, submerged the seeds in the water, removed from heat, placed a lid on the pot and let sit for 40ish minutes.
4. Rinsed the seeds in a strainer until water was clear, set on the counter with a fan to dry for 2 hours, then separated 850g~ into each mason jar.
5. Covered the lids in foil with a rubber band and placed in PC.
6. Ran the PC for 10 minutes to vent then 90 minutes at 15psi.

Photos









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InvisibleMicroMycelium
Curious Optimizer

Registered: 06/06/23
Posts: 461
Loc: Probably at work
My First Agar Brew [Re: MicroMycelium] * 1
    #28362770 - 06/16/23 11:22 PM (7 months, 8 days ago)

Ordered some 90ml glass plates, sterilized 15 minutes at 15psi, let cool then filled with my new agar brew.

MEPYAC Agar:
This is Malt/Peptone/Yeast/Charcoal Agar.

Recipe:
- 500ml distilled water
- 9g Agar
- 7g Light Malt Extract
- 0.5g Peptone
- 0.5g Nutritional Yeast
- 1g Activated Charcoal

Process:
1. Add ingredients to 100ml distilled water in media bottle
2. Stir with electronic mixer & magnetic pill for a few minutes
3. Pour 400ml of boiling distilled water into media bottle while still mixing, continue mixing for 3-5 minutes.
4. Place lid on bottle place bottle into PC. 15psi for 20 minutes.
5. Remove the bottle and let cool until roughly 120 degrees.
6. Within my new still air box, poured agar into newly sterilized glass plates.
7. Secured plates into a 6qt box for future use :thumbup:

Photos:





Edited by MicroMycelium (06/16/23 11:23 PM)


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OfflineExbat
New Oldie
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Registered: 10/03/22
Posts: 44
Loc: PNW, U.S.A.
Last seen: 6 months, 24 days
Re: WBS Grain Spawn [Re: MicroMycelium] * 1
    #28362881 - 06/17/23 04:16 AM (7 months, 7 days ago)

Quote:

MicroMycelium said:
WBS grain spawn

-1qt Mason jars with no-leak lids
-Roughly 850g WBS each

Process

1. Put all the grains in a pot and sifted/rinsed with cold water 7-10 times until the water was relatively clear.
2. Pulled 99% of the sunflower seeds floating on the water out.
3. Brought a pot of water to a boil, submerged the seeds in the water, removed from heat, placed a lid on the pot and let sit for 40ish minutes.
4. Rinsed the seeds in a strainer until water was clear, set on the counter with a fan to dry for 2 hours, then separated 850g~ into each mason jar.
5. Covered the lids in foil with a rubber band and placed in PC.
6. Ran the PC for 10 minutes to vent then 90 minutes at 15psi.

Photos












I follow this admittedly dated WBS tek and have never dried the WBS after soaking. After draining the grains may be somewhat damp still but that is absorbed during PCing. Shake the jars after PCing to distribute dampness and all is well.


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InvisibleMicroMycelium
Curious Optimizer

Registered: 06/06/23
Posts: 461
Loc: Probably at work
Re: WBS Grain Spawn [Re: Exbat] * 1
    #28362886 - 06/17/23 04:27 AM (7 months, 7 days ago)

That gives me peace of mind, because there was a decent amount of condensation in the jars after the PC lol


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InvisibleMicroMycelium
Curious Optimizer

Registered: 06/06/23
Posts: 461
Loc: Probably at work
Germ Plate [Re: MicroMycelium] * 1
    #28363625 - 06/17/23 06:54 PM (7 months, 7 days ago)

Decided to put my fully colonized germ plate from one of my MSS into a grain jar.

I figured since the plate was fully colonized and I had already taken the sectors I wanted I may as well throw it into one of my grain jars.

I also took another sector to test on my new black agar.


Assuming I did a decent job with my grains, it should be interesting to see all the different phenotypes compete to colonize it.


The Germ plate looks contam-free right?




Edited by MicroMycelium (06/17/23 06:55 PM)


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Offlinetripdawg420
low life with no life
Male


Registered: 02/02/09
Posts: 7,071
Loc: illinois
Last seen: 7 hours, 10 minutes
Re: Germ Plate [Re: MicroMycelium] * 1
    #28363648 - 06/17/23 07:23 PM (7 months, 7 days ago)

:takingnotes:


--------------------
HUSTLER
How U Survive This Life Everyday Resourcefully
epic GT mono tub
http://www.shroomery.org/forums/showflat.php/Number/17277772

wbs tek
http://www.shroomery.org/forums/showflat.php/Number/11525679
coir tek
http://www.shroomery.org/forums/showflat.php/Number/11917410
results :thumbup:


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InvisibleMicroMycelium
Curious Optimizer

Registered: 06/06/23
Posts: 461
Loc: Probably at work
New germ plate [Re: MicroMycelium] * 1
    #28365163 - 06/19/23 07:19 AM (7 months, 5 days ago)

I was looking at my T2 transfers to my new agar plates and noticed the mycelium immediately took to the MEPYAC agar formula I mixed better than the premade PDA plates I bought.

Naturally, I put one drop from my first MSS syringe on it to test how spores take to it comparatively.
Hoping to see some faster germination.



Edited by MicroMycelium (06/19/23 08:12 PM)


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InvisibleMicroMycelium
Curious Optimizer

Registered: 06/06/23
Posts: 461
Loc: Probably at work
Agar & Grain update [Re: MicroMycelium] * 1
    #28366228 - 06/19/23 08:12 PM (7 months, 5 days ago)

Update on my T1 & T2 plates and grain jar with germ plate.


Here are my original T1 transfers on PDA plates.
Original germ plate was inoculated on 5/29 with a tidal wave MSS.
That makes these now 21 days/3 weeks in. 2 weeks from being transferred




It's pretty cool to see the difference in mycelium growth rates & textures from the different phenotypes.


These are my T2 transfers on my MEPYAC plates from 06/18.
That makes these almost 48hrs old.
(Top left is another drop from the MSS on 06/18 midnight to compare germination rate/quality on different agar formula)



Here is my WBS quart grain spawn jar with the original MSS spore plate thrown in.
Approximately 48hr in and already visible mycelium spreading to grain.



Edited by MicroMycelium (06/19/23 08:15 PM)


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OfflineTheLiesUnderneath
Lost Prophet
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Registered: 05/19/16
Posts: 222
Last seen: 1 month, 2 days
Re: Agar & Grain update [Re: MicroMycelium] * 1
    #28366432 - 06/19/23 11:39 PM (7 months, 5 days ago)

Good job, it's been so long since I've grown. This grow log is great inspiration. Thank you my friend


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InvisibleMicroMycelium
Curious Optimizer

Registered: 06/06/23
Posts: 461
Loc: Probably at work
Update [Re: MicroMycelium] * 1
    #28373468 - 06/24/23 10:23 PM (7 months, 4 hours ago)

Spent the last weekend hiking 9 miles in Utah's national park Zion on 1g lemon tek PE. Highly recommend. Here's an update I came back to.

Here is my first grain jar made from my MSS Germ plate on 6-17, so now 7 days in.
Looks clean to me, keep in mind it's a germ plate so I don't expect very "uniform" mycelium.




Here are some random empty Agar plates that must of caught some contams when I poured them in my SAB.
I also put some rehydrated PE in the top plates... total failure lol. Will be cleaning these out and re-sterilzing them at some point.


Here are my MEPYAC agar transfers and MSS plate from 6-18 so now 6 days in.
So far so good, however growth actually looks slower than my PDA plates.


Here are my original PDA plates from 6/07 & 6/10. A couple looked good and colonized their entire plates so I took a sector from the best "grain" marked plates, and threw in my second germ plate to another jar.



And the Jars:

Note I put one plate in a jar with a lid that has Sterile Filter Disks to compare to just cracking my no-leak lid on the first.


Edited by MicroMycelium (06/24/23 10:26 PM)


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InvisibleMicroMycelium
Curious Optimizer

Registered: 06/06/23
Posts: 461
Loc: Probably at work
Agar variations [Re: MicroMycelium] * 1
    #28376481 - 06/27/23 11:43 AM (6 months, 28 days ago)

My new agar is growing much different than my PDA plates.

One plate, which looks clean, has formed moisture central to its transfer a week after being transferred from a T1 plate.



The other plates are developing at a decent rate however 90% of them have a sort of dry halo that traces around the mycelium.




My theory is that maybe because the agar has more nutrition it rapidly expanded and will thicken up as time goes on..?

If anyone has seen this stuff before feedback is appreciated


Edited by MicroMycelium (06/27/23 11:47 AM)


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InvisibleMicroMycelium
Curious Optimizer

Registered: 06/06/23
Posts: 461
Loc: Probably at work
Glass Petri dishes no go [Re: MicroMycelium] * 1
    #28378020 - 06/28/23 04:57 PM (6 months, 27 days ago)

Never again will I buy autoclavable glass Petris.
These are the ones I got, the 90mm.
https://a.co/d/isaTE9y

The lids that come with have a significant gap between the base. So much that after PC at 15psi, pouring in my SAB, and then storing in a closed sanitized drawer which is housed in a sealed grow tent, 7 out of 18(two broke during shipping) all have contaminants in them.

After noticing two of them being contaminated the day after pouring,  I’ve checked the blank ones every few days and each time I’ve checked a new dish has something growing on it.

I assume this is from handling them since the lid doesn’t sit flush and allows air to get in just picking them up.

The only reliable way to use these is to probably put parafilm on every single one even when not in use because even dish I put a transfer in and sealed with parafilm is clean… which shows the non-sealed are vulnerable just existing


Edited by MicroMycelium (06/28/23 04:59 PM)


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Offlinebongoman
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Registered: 09/19/18
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Re: Glass Petri dishes no go [Re: MicroMycelium] * 1
    #28378078 - 06/28/23 05:49 PM (6 months, 27 days ago)

I use glass plates and always wrap them after they've cooled in front of the hood. But I did that when pouring plastic plates too.


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InvisibleMicroMycelium
Curious Optimizer

Registered: 06/06/23
Posts: 461
Loc: Probably at work
Attempted to save the a few contaminated dishes [Re: MicroMycelium] * 1
    #28381568 - 07/01/23 04:25 PM (6 months, 24 days ago)

I flame sterilized my scalpel and cut around the bacteria spots I saw form on two of my new plates in my SAB in attempt to save them.

I did this to 3 blank plates and two plates with cultures on them.
Also threw two more of my PDA plates to grain and shook the previous three.

To my inexperienced eye the grain jars look good so far, maybe a little dry since I didn't soak my WBS but clean and the mycelium definitely is spreading.
I'll get some photos up tomorrow or the day after.


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InvisibleMicroMycelium
Curious Optimizer

Registered: 06/06/23
Posts: 461
Loc: Probably at work
Waiting game [Re: MicroMycelium] * 1
    #28383416 - 07/03/23 09:54 AM (6 months, 22 days ago)

Update:
Just got a 30qt bag of Verm, 3lb bag of gypsum, and some new LC lids(which I’ll use from a completely rhizomorphic plate if I ever get one).

I’ve got 6 WBS jars colonizing each between 20-50% ready.
Just patiently… waiting… patiently.

Waiting sucks and I want to try and spawn my jars :frown:


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