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Brazilian Cubensis grow log Options
 
dmusicaltrancistor
#1 Posted : 11/21/2017 8:58:32 PM

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It has been a couple years since the last time I had the opportunity to work with some Cubensis and the time has arrived so I figured why not share my work as I go along.

In the past I have been blessed with a few different great teachers whom have been gracious enough to impart their knowledge and artistry of growing mushrooms.

So to start a friend and I have decided to start a Brazilian Psilocybe Cubensis grow we will start with a multi spore syringe and cover a few different methods of growing.
1. A batch of MS BRF cakes
2. A MS Liquid Culture (LC)
3. A MS agar plate strain isolation
4. A clone to agar strain isolation

The point of this experiment is to show the method the I have found to be most useful in a. Getting started growing mushrooms for a first timer. B. Quickly and efficiently building your genetics to ensure strong future genetics and prevent strain degradation. I also want to outline the different approaches people take to growing mushrooms and why/when different approaches are used.
dmusicaltrancistor attached the following image(s):
Glove Box.JPG (2,249kb) downloaded 174 time(s).
Spore Syringe.JPG (1,793kb) downloaded 174 time(s).
Cubensis Mushroom Colonization Temps 75-80 F Cubensis Mushroom Fruiting Temps 68-74 F
https://ocw.mit.edu/reso...ues-manual-spring-2007/
https://www.dmt-nexus.me/forum/default.aspx?g=posts&t=36239
cyb wrote:
Xtals are just goo with fancy designer clothes on...
Big grin

 

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dmusicaltrancistor
#2 Posted : 11/21/2017 9:45:00 PM

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So i would like to mention my photos are on my phone and will be uploaded shortly

So my Friend (from here on we will call zen1) and I put our alchemical powers together and conjured up some brazillian cubensis spores
on Nov 1st we then proceeded to mix and sterilize 5 BRF cakes following the ever popular PF tek utlined in this video series
https://www.youtube.com/watch?v=ZHJQrsZFQdE

After we mix and sterillized our jars they were then innoculated with approximately .5cc of fluid from the spore syringe
The jars were then placed in a incubation chamber with a temperature set around 30c and checked on occasionally over the course of the next few weeks while waiting for the jars to colonize with mycellium


My incubation chamber consists of a 3 inch deep plastic tray filled half way with water. Under the tray i have placed a heating blanket with multiple heat settings to ensure the correct temperature is acheivable.
inside of the tray with water i have placed my PF jars and then covered the tray with a piece of cardboard that has a thermometer stuck through to monitor water temp
dmusicaltrancistor attached the following image(s):
IMG-0192.JPG (1,499kb) downloaded 178 time(s).
Cubensis Mushroom Colonization Temps 75-80 F Cubensis Mushroom Fruiting Temps 68-74 F
https://ocw.mit.edu/reso...ues-manual-spring-2007/
https://www.dmt-nexus.me/forum/default.aspx?g=posts&t=36239
cyb wrote:
Xtals are just goo with fancy designer clothes on...
Big grin

 
dmusicaltrancistor
#3 Posted : 11/21/2017 10:18:29 PM

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Zen1 and Myself then decided a Multi Spore Liquid Culture was definately called for it being Zen1's first proper mushroom grow we thought it a good idea to cover all bases so after we had let our MS PF jar get their orgy on for a couple weeks on Nov 15th we started our Brazillian Multi Spore Liquid Culture following the easy microwave Karo Tek.
A quick outline of this is

take 100ml distilled water add 4 grams of Karo (or light corn syrup) place in microwave for 30 secs to warm up karo/water then mix and and place back in microwave for 5 mins imediately take out with oven gloves and screw on jar lid with filter disc and self sealing innoculation port wait until room temp then innoculate with spore syringe

after finishing this procedure the jar was then placed in the incubation chamber with the other PF jars

PF jars were check for mycellium growth at this time and we noticed the first signs of growth in 2/5 Jars

all jars were then placed back in incubating chamber and kept at a constant 30c
dmusicaltrancistor attached the following image(s):
IMG-0204.JPG (1,316kb) downloaded 177 time(s).
Cubensis Mushroom Colonization Temps 75-80 F Cubensis Mushroom Fruiting Temps 68-74 F
https://ocw.mit.edu/reso...ues-manual-spring-2007/
https://www.dmt-nexus.me/forum/default.aspx?g=posts&t=36239
cyb wrote:
Xtals are just goo with fancy designer clothes on...
Big grin

 
dmusicaltrancistor
#4 Posted : 11/21/2017 10:32:08 PM

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So at this point it is now Nov 21st All of our PF jars have healthy looking growth in them with no signs of contamination Thumbs up

our LC has definate strands of mycellium growing and looks clear not cloudy hopefully signifying no contams Thumbs up


So far so good

we are currently waiting on an order of petri dishes to start a MS strain issolations and as soon as we have some fruits we will cover; 1. Harvesting times
2. Spore Printing
3. Fruit choices for cloning
4. cloning to agar
5. cloning to LC

dmusicaltrancistor attached the following image(s):
All Jars.JPG (1,318kb) downloaded 176 time(s).
Jar 1.JPG (937kb) downloaded 176 time(s).
Jar1 Side 2.JPG (1,021kb) downloaded 176 time(s).
Jar2.JPG (1,046kb) downloaded 176 time(s).
Jar 2 Side 2.JPG (1,119kb) downloaded 175 time(s).
jar3.JPG (1,721kb) downloaded 175 time(s).
Jar3 Side 2.JPG (1,142kb) downloaded 174 time(s).
Jar4.JPG (1,684kb) downloaded 175 time(s).
Jar4 Side 2.JPG (1,089kb) downloaded 174 time(s).
Jar5.JPG (1,388kb) downloaded 175 time(s).
Jar5 Side 2.JPG (986kb) downloaded 175 time(s).
Liquid Culture.JPG (1,316kb) downloaded 175 time(s).
Cubensis Mushroom Colonization Temps 75-80 F Cubensis Mushroom Fruiting Temps 68-74 F
https://ocw.mit.edu/reso...ues-manual-spring-2007/
https://www.dmt-nexus.me/forum/default.aspx?g=posts&t=36239
cyb wrote:
Xtals are just goo with fancy designer clothes on...
Big grin

 
dmusicaltrancistor
#5 Posted : 11/25/2017 5:15:42 AM

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Grow log update Nov 24

Today we are starting MS strain isolation on agar from spore syringe and WBS bulk grain spawn from spore syringe, tomorrow we will start MS LC strain isolation and MS LC WBS bulk grain spawn.

The agar was made by mixing 4g fish food flakes with 4g agar agar in a medium jar then dissolving with 200 ml boiling water the jar is the sealed and steam sterilaized for 45 mins and allowed to partially cool. 3 jars are then sterillized and filled approximately 1 cm with agar mixture these are then sealed and allowed to finish cooling. The jars are the inoculated with 1 drop each from the spore syringe. At this point the jars are sealed and placed into colonization conditions to allow mycellium to begin to grow.

as i has mentioned we are also starting 3 WBS bulk grain spawn jars so i can quickly go over what has been done so far.
We filled 3 jars 1/3 of the way up with WBS then added water until it came up to the 2/3 mark we then stirred the grains to allow all of the black sunflower seeds to float up and removed them. At this point the jars had the lids losely placed on them and have been set aside to soak for the next 12 hours.

pictures are of the 3 inoculated agar jars, The 3 WBS grain spawn jars, and the removed black sunflower seeds
dmusicaltrancistor attached the following image(s):
Agar Jars.jpg (1,166kb) downloaded 157 time(s).
WBS grain spawn jars.jpg (1,087kb) downloaded 157 time(s).
BS seeds.jpg (1,444kb) downloaded 156 time(s).
Cubensis Mushroom Colonization Temps 75-80 F Cubensis Mushroom Fruiting Temps 68-74 F
https://ocw.mit.edu/reso...ues-manual-spring-2007/
https://www.dmt-nexus.me/forum/default.aspx?g=posts&t=36239
cyb wrote:
Xtals are just goo with fancy designer clothes on...
Big grin

 
tatt
#6 Posted : 11/25/2017 10:36:03 AM
[tôt]

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Looking great dmusicaltrancistor Smile


Thanks for documenting all this, such a beautiful process; never gets old to watch them slowly but surely colonize over the time span. Love
For to be idle is to become a stranger unto the seasons, and to step out of lifes procession, that marches in majesty in proud submission towards the infinite. - Khalil Gibran



 
dmusicaltrancistor
#7 Posted : 11/26/2017 4:44:40 PM

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^^^ Thanks Tatt much love goes out to you

Sorry if anyone was waiting on this update but I didn't have time to get online yesterday but here is what we accomplished.

Yesterday Nov 25th
The 3 Jars previously mixed with WBS and water had been left to soak for 12-16 hrs at which point they were strained and rinsed with clean water. All of the WBS was then combined into 1 pot and 2 qts of water was added. This was brought to a boil and allowed to lightly boil with the lid on for 45 mins. The WBS was then strained and rinsed with clean boiling water then spread out on paper towel for quick steam drying through evaporation. 3 qt jars were then filled 2/3 of the way with the prepared WBS lids were placed on the jars then covered with tin foil. The jars were then steam sterilized for 90 mins and allowed to cool to room temperature in the pot with the lid still on. After cooling jars are removed from the pot and inoculated with 1-2cc each from the Brazil MS Spore syringe and then placed into colonization conditions.

Awesome so... after all that was finished we started soaking another 3 jars of WBS that we will be sterilizing today using the same method listed above. These will be inoculated with a Liquid Culture we had made with the MS Spore syringe 11 days ago. By having 3 jars inoculated via MS Spore Syringe and another 3 jars inoculated via MS Liquid Culture we hope to demonstrate the difference in colonization times between the 2 methods. Even though the jars are "Noc'd up" a day apart we should still be able to observe a difference.
After we have finished this I will update again and with pics.
Cubensis Mushroom Colonization Temps 75-80 F Cubensis Mushroom Fruiting Temps 68-74 F
https://ocw.mit.edu/reso...ues-manual-spring-2007/
https://www.dmt-nexus.me/forum/default.aspx?g=posts&t=36239
cyb wrote:
Xtals are just goo with fancy designer clothes on...
Big grin

 
downwardsfromzero
#8 Posted : 11/26/2017 5:30:20 PM

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Great that you're doing this. I eagerly await comparison of the results from WBS vs PF cakes. You probably know, the WBS will rock the joint Thumbs up
All posts by this user are purely fictitious and provided for entertainment purposes only.
Ora, lege, lege, lege, relege et labora
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Thinking of evaporating? Save money and the environment! Reuse your solvents!
 
dmusicaltrancistor
#9 Posted : 11/26/2017 6:30:40 PM

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Ya I am expecting them to colonize much quicker than the BRF cakes and I expect that the 3 jars Noc'd With the LC to Colonize much Quicker than the 3 jars Noc'd with the MS Syringe. As I go I am collecting all of the time it takes for each method and final weights for each as well. I will also cover strain isolation to LC to grain and also the benefits of using different additives to your grains, bulk substrate, Casing layer.
a few of those additives will be: Dilute Coffee
Gypsum
Lyme
Crushed oyster shells

At the end of everything my goal is to have a consolidated list of everything above, so anyone can easily see the difference of each method with data instead of opinion.

Hopefully anyways but either way it should help anyone interested in the cultivation process wether they are just starting out or looking to try a new technique like Bulk Growing, Strain Isolation, etc.
Cubensis Mushroom Colonization Temps 75-80 F Cubensis Mushroom Fruiting Temps 68-74 F
https://ocw.mit.edu/reso...ues-manual-spring-2007/
https://www.dmt-nexus.me/forum/default.aspx?g=posts&t=36239
cyb wrote:
Xtals are just goo with fancy designer clothes on...
Big grin

 
dmusicaltrancistor
#10 Posted : 11/27/2017 4:34:38 PM

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Update for yesterday Nov 26

We started the day by sterilizing a new syringe and filling it with 10cc from the Liquid Culture we had started on Nov 15th inside of our still air box, then set it aside to be used later on.

The 3 Jars previously mixed with WBS and water had been left to soak for 12-16 hrs at which point they were strained and rinsed with clean water. All of the WBS was then combined into 1 pot and 2 qts of water was added. This was brought to a boil and allowed to lightly boil with the lid on for 45 mins. The WBS was then strained and rinsed with clean boiling water then spread out on paper towel for quick steam drying through evaporation. 3 qt jars were then filled 2/3 of the way with the prepared WBS lids were placed on the jars then covered with tin foil. The jars were then steam sterilized for 90 mins and allowed to cool to room temperature in the pot with the lid still on. After cooling jars are removed from the pot and inoculated with 1-2cc each from the MS Liquid Culture Syringe and then placed into colonization conditions.

While we were prepping the grains we also made more agar and agar plates the agar was made by boiling 400 ml water then adding 8g Fish Food Flakes and 8g Agar-Agar this was mixed and then poured into a quart jar. Some of the prepared agar was used to make another 2 agar plates, these 2 plates will be used later for strain isolation. The agar Jar and Agar plates were then steam sterilized for 90 mins along with the 3 WBS grain Jars.
Both plates were allowed to cool after sterilization and then Noc'd with 1-2 drops each from the MS Liquid Culture Syringe, then placed into colonization conditions.
The excess Agar jar was placed in the fridge for future use.

and a bonus not related to the grow we also brewed up 10g of syrian rue tea for a Pharma journey soon to be had.
Cubensis Mushroom Colonization Temps 75-80 F Cubensis Mushroom Fruiting Temps 68-74 F
https://ocw.mit.edu/reso...ues-manual-spring-2007/
https://www.dmt-nexus.me/forum/default.aspx?g=posts&t=36239
cyb wrote:
Xtals are just goo with fancy designer clothes on...
Big grin

 
dmusicaltrancistor
#11 Posted : 12/1/2017 4:41:32 PM

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Update Dec 1st

Figured I would give you guys a quick update My agar plates are growing out nicely, the BRF cakes have no contams and are sitting anywhere from 60-95% colonized, and the WBS jars are starting to show some signs of mycelium
Cubensis Mushroom Colonization Temps 75-80 F Cubensis Mushroom Fruiting Temps 68-74 F
https://ocw.mit.edu/reso...ues-manual-spring-2007/
https://www.dmt-nexus.me/forum/default.aspx?g=posts&t=36239
cyb wrote:
Xtals are just goo with fancy designer clothes on...
Big grin

 
Wakinyan
#12 Posted : 12/1/2017 5:03:53 PM

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Have you done any spore isolation via serial dilution and plating? Growing out on agar plates to mate individual strains with other strains in a more controlled manner allows for controlled mating and isolation....
When I graft you graft we graft
 
dmusicaltrancistor
#13 Posted : 12/1/2017 5:50:30 PM

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Wakinyan wrote:
Have you done any spore isolation via serial dilution and plating? Growing out on agar plates to mate individual strains with other strains in a more controlled manner allows for controlled mating and isolation....


No I havent but you have peaked my interest can you explain the process to me i think i understand the basics but would love to know more then when I start my next round of plates from spore i will give it a shot
Cubensis Mushroom Colonization Temps 75-80 F Cubensis Mushroom Fruiting Temps 68-74 F
https://ocw.mit.edu/reso...ues-manual-spring-2007/
https://www.dmt-nexus.me/forum/default.aspx?g=posts&t=36239
cyb wrote:
Xtals are just goo with fancy designer clothes on...
Big grin

 
Wakinyan
#14 Posted : 12/1/2017 6:36:08 PM

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It is very simple. You set up a serial dilution via test tubes normally, but any container will work provided it is sterile.

Label your first container 1 and fill this up with a known working dilution of spores you would normally inoculate with.

Label your second container 2 and fill this container half as full of sterile water as your previous container. Take half the spore dilution from container one and now place that in container 2. Make sure to mix each time.

Label your third container 4 and again fill this container up halfway with sterile water. Pull 1/2 of the spore dilution from container 2 and mix into your container labeled 4.

next container 8 then 16 then 32, 64, etc.

Once you have made these dilutions you will simply take one drop from each dilution and plate that onto an agar plate you have made. I get the agar with a bit of antibiotics added into them just in case and find it is worth the extra step.

That one drop can then be mixed via a 3 or 4 quadrant streak if your familiar with this practice as it is taught in most first year microbiology courses, but it can also be found online. It takes some practice, but you should be proficient enough to isolate a single spore colony in quadrant 3 or 4 in a few tries even if you have never done it before. The main thing is to never breath on your plates as that is a sure fire way of introducing microorganisms if any are floating about. Also, label your plates according to the dilution you used and then flip them upside down to incubate after they have sat long enough to absorb that one drop... usually less than 30 seconds to 1 minute so by the time you finish your plates you will be good to go. Flipping them upside down also keeps the moisture from condensing and falling on your spores further exasperating your problem. Once you have a single spore isolated you can then take that spore and transfer it to its own plate to keep it pure. Whenever you wish to cross another spore you will simply dip your inoculation loop into the pure culture and mix dab it onto the right side of a plate. Label that inoculum with whatever number or name you have given to your spore to differentiate it from other spores from that same strain. Next, simply plate or inoculate the opposite side with another spore from a different strain or even the same strain if you are looking to improve that strain or hybridize... you get the basic idea and can go from there. Always keep your spores separate so you can test which ones work best together and most importantly either flame your loop between inoculations or use a disposable loop. When in doubt, chuck it out. You never want to contaminate your mother cultures.
Hope that helps
When I graft you graft we graft
 
Wakinyan
#15 Posted : 12/1/2017 6:39:06 PM

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One ore quick addition, if you aren't familiar with a 4 quadrant streak or have shaky hands you can always use a sterile glass bar and roll that through your drop to spread your spore dilution around. It is not quite as good, but once you get a dilution down to where you only have 1-10 spores per drop you really can't mess it up and should be able to find many single spore colonies at any rate. With good technique however, you won't even need to dilute... just streak away and you will most assuredly get some isolated colonies.
When I graft you graft we graft
 
dmusicaltrancistor
#16 Posted : 12/6/2017 4:33:00 PM

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Holy DiMiTri I has been busy Pleased The run of BRF cakes have finished colonizing I placed the first one finished into a mini Dub Tub made out of a couple ziplock containers, so that i can fruit and get some nice spore prints and some clone tissue for agar work.
The other 4 cakes were all used for spawning to bulk each given their own mono tubs this was done over the course of Dec 4th to Dec 5th.

We also got 2 more varieties of spore syringe; Ecuadorian P.C., and PES Amazonian P.C. so I made 2 BRF jars, and 1 Master WBS spawn jar of each of these new strains.
-The BRF cakes will be used to gather spore prints (for future use and trades) and clone tissue (for agar work)
-The WBS jars will be used for master spawn to make more WBS jars

Dec 5th.
After making my cakes and jars I had left over BRF mix which was put in a oven bag to be sterilized with the BRF cakes and I had a bunch of leftover coir which was place in oven an bag to be sterilized. I also still had some of my LC left so I used that to Noc up the bag of BRF and figured i would also try straight LC to coir as well so put the coir in a mono tub and poured the remaining LC over top as an experiment which I think probably has less than 50% chance of working but I was going to toss the leftover materials so why not try and see what happens.
And Moar LC... Laughing seeing as we got some new spore syringes why not expand the inoculation potential by making more LC so we whipped up 2 400ml batches of water and karo sterilized and Noc'd up 1 jar with 1cc of the Ecuadorian and 1 jar with 1cc of the PES Amazonian.

so to review
Dec 3rd - 1 BRF jar was birthed and dunked
- 2 BRF jars were Birthed and used to spawn 2 bulk mono tubs (No coffee or gypsum)
Dec 4th - Dunked BRF jar was placed in mini Dub Tub
Dec 5th - 2 BRF jars were Birthed and used to spawn 2 bulk mono tubs (coffee and gypsum)
- 2 WBS jars were made and Noc'd with new strains (coffee and gypsum)
- 4 BRF jars were made and Noc'd with new strains (coffee and gypsum)
- 2 LC jars were made and Noc'd with new strains
- 1 BRF bag was made and Noc'd with Brazilian LC (coffee and gypsum)
- 1 Mono Tub was made and Noc'd with remaining Brazilian LC (coffee and gypsum)

The Mono tubs spawned on Dec 3rd are already 25-35% colonized and the BRF in the Dub Tub is showing some pins starting as well
Big grin Love


P.S. forgot to mention dilute coffee water and gypsum were used in the BRF mix the WBS soak and simmer, and the coco coir/Verm Bulk substrate dilute coffee was used in a 50/50 ratio with water in each step requiring water and a small handful of gypsum was added during the mix process of the BRF, added in the mixing process of the bulk substrate, and in the soak and simmer step of the WBS
Cubensis Mushroom Colonization Temps 75-80 F Cubensis Mushroom Fruiting Temps 68-74 F
https://ocw.mit.edu/reso...ues-manual-spring-2007/
https://www.dmt-nexus.me/forum/default.aspx?g=posts&t=36239
cyb wrote:
Xtals are just goo with fancy designer clothes on...
Big grin

 
dmusicaltrancistor
#17 Posted : 12/6/2017 9:17:18 PM

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Well after all that work my buddy wakes up this morning and surprises me with yet another strain of cubes to work with (Really wishing he had of done this before I did everything yesterday) Soooo it looks like I have some moar work to do... I shall be back with moar updates soon when I get a chance
Cubensis Mushroom Colonization Temps 75-80 F Cubensis Mushroom Fruiting Temps 68-74 F
https://ocw.mit.edu/reso...ues-manual-spring-2007/
https://www.dmt-nexus.me/forum/default.aspx?g=posts&t=36239
cyb wrote:
Xtals are just goo with fancy designer clothes on...
Big grin

 
dmusicaltrancistor
#18 Posted : 12/9/2017 1:56:51 AM

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Pics of the growing stack of BRF jars, WBS jars, and Monotubs
and... a bonus xtal shot
dmusicaltrancistor attached the following image(s):
5 Mono's.jpg (1,583kb) downloaded 42 time(s).
BRF and WBS Colonizing.jpg (1,579kb) downloaded 42 time(s).
LC + MiniMono.jpg (1,480kb) downloaded 42 time(s).
xtals.jpg (1,163kb) downloaded 42 time(s).
Cubensis Mushroom Colonization Temps 75-80 F Cubensis Mushroom Fruiting Temps 68-74 F
https://ocw.mit.edu/reso...ues-manual-spring-2007/
https://www.dmt-nexus.me/forum/default.aspx?g=posts&t=36239
cyb wrote:
Xtals are just goo with fancy designer clothes on...
Big grin

 
dmusicaltrancistor
#19 Posted : 12/10/2017 6:14:25 AM

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Noc'd up 4 WBS jars today with a new strain this time its texas apparently this has two variations one with an orange cap and one with a yellow cap so well see if i can isolate out these traits and have a yellow agar plate and orange agar plate.... and also the yellows will sometimes throw a leucistic fruit so hopefully that will happen one day and i can work towards stabilizing an albino texan if im really stupid lucky
Cubensis Mushroom Colonization Temps 75-80 F Cubensis Mushroom Fruiting Temps 68-74 F
https://ocw.mit.edu/reso...ues-manual-spring-2007/
https://www.dmt-nexus.me/forum/default.aspx?g=posts&t=36239
cyb wrote:
Xtals are just goo with fancy designer clothes on...
Big grin

 
dmusicaltrancistor
#20 Posted : 12/10/2017 6:40:36 PM

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Dec 10th 2017

We have reached pinspermia i got some massive looking pin heads on the Brazil BRF cake right now cant wait another couple days and i will have some nice fresh spore prints from this bad boy and hopefully some prime clone tissueTwisted Evil Love
Cubensis Mushroom Colonization Temps 75-80 F Cubensis Mushroom Fruiting Temps 68-74 F
https://ocw.mit.edu/reso...ues-manual-spring-2007/
https://www.dmt-nexus.me/forum/default.aspx?g=posts&t=36239
cyb wrote:
Xtals are just goo with fancy designer clothes on...
Big grin

 
 
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