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Another look at Psilocybe Cubensis cultivation Options
 
Espurrr
#1 Posted : 7/3/2019 2:08:41 AM




Posts: 403
Joined: 23-Aug-2015
Last visit: 09-Mar-2024
Location: Iran

spore syringe:

using spore syringes are relatively simple, i would just use them to inoculate 0.5kg spawn jars or bags and fruit them in a smaller box to sample tissue from the fruit stems on agar, but you can just continue taking spore prints from fruits and making more spore syringes and grow your mushrooms that way and it works, just stick with 0.5 to 1 kg spawn and colonize larger spawn bags with master grain (aka smaller colonized spawn) to prevent contamination

Agar:

1- Bring 1 liter of reverse osmosis or spring water to a boil

2- Add 30g light malt extract (you could also use filtered grain water from your spawn cook) + 3g nutritional yeast extract + 2g activated carbon (optional but makes for easier contamination recognition)

3- Slowly add 30g of agar/liter and stir until fully dissolved

4- Pour the agar medium to fill half of autoclave glass bottles (has a plastic screwed lid) or mason jars with a cotton filled hole and sterilize for 30 minutes at 15psi or 45 minutes in a home pressure cooker (open the de-pressure valve at a minimum and when the pressure inside your unit is the same as the room proceed to take the glasses out, fill 75% the volume of the glass MAX to prevent overflow)

5- Turn on your laminar flowhood about 30 minutes before pouring the agar in plates if it doesn’t come with an enclosed glass box to work in (the room should be closed off with positive pressure airflow from a hepafilter), Wear latex gloves and a breathing mask and sterilize your hands with 70% isopropyl-ethanol and lysol to ensure sterility

Let the solution cool to 60 Celsius and pour into agar plates in front of your laminar flow hood or Still Air Box, make sure to pour enough so that you fill half or more of the plate volume

6- Store the freshly poured plates in a clean wiped (with lysol and 70% isopropyl) box if your flowhood doesn't come with a glass box to ensure sterility while the plates cool and get ready for transfers

7- Use a scalpel and wipe it down with 75% isopropyl and flame sterilize it with a butane torch after the isopropyl evaporates (unless the blades come already sterilized) and let it cool before using it to transfer tissue from another plate or fruiting body or spore print

While transferring choose the healthy and rigorous sector of the mycelium and wrap the agar plate closed with breathable parafilm or micropore tape or other suitable materials to prevent contamination and water loss from the media

Colonize in 23C-27C (possible up to 30c but that stresses the mycelium over time and makes for more contaminations etc) and store the plates upside down in the fridge after

Liquid culture:

LC jars need a 2 micron filter patch or cotton filled holes and a silicone syringe port which you can make by using heat resistant silicone adhesives or just open the lid if you have a flowhood (make sure not to dent or bend lids and make perfect holes and ) and magnet bars (~3cm), make sure your jar or whatever you use has a properly fitting lid


Use 2g of light malt extract in 1L of reverse osmosis or spring water, fill about half of the jars (make sure to clean the jars if they were used with 20% bleach or lysol and isopropyl before using) with a magnetic bar (3cm) included and autoclave for 30 minutes at 15psi and colonize at 23C-27C and stir on fast on the magnetic stirrer for until tissue shreds everyday, use the lc or store in the fridge in a clean box before it gets too thick to suck up with a syringe, otherwise you can just pour it in your spawn instead of using a syringe

Autoclave the syringe and needle for 30m (unless they come already sterilized) and cool it in front of the flowhood/ in the still air box (i use a needle with a wider diameter to suck up a thicker LC as that helps alot with better colonization) and suck up the liquid culture and close the needle port and wrap with cellophane if you want to store it in the fridge for later, can also wrap the LC jar and store it in the fridge for later

Spawn (probably the most sensitive stage):

-i soak and rinse and strain with no cooking using organic millets with spring or tap water that's been sitting for a day for spawn bags (rinse completely until all dirt is gone before using) 1000g soaked millets + 1g gypsum in each spawn bag after my millets have absorbed all the water they can which usually takes less than 24h, i rinse to clean and strain them until all the extra water is gone and put them in spawn bags to sterilize in the autoclave, if i want to inject spores or liquid cultures or put in agar plates aka make master spawn bags, and then use 2kg and 3kg every time i do spawn to spawn in the bags to expand for more colonized spawn, this is the soak and no cook method with added gypsum after straining which has worked wonders for me

push all the air out if you're using spawn bags and wrap the bag over the spawn so it doesn't open back up before autoclaving (which vacuum seals your spawn automatically)
autoclave at 15 psi for 90 mins for 1 kg to 180 mins for 2 kg depending on the size of your spawn bags

Inject each 1kg spawn bag with about 3ml of thick LC in front of the flow hood (wipe the injection site with 70% iso and lysol or use a self healing injection port, you can also just open the top and pour in liquid cultures or agar wedges, tho if using spore syringe don't shake or move too much after injection to keep the spores in one place)

Colonize at 23C-27C and shake twice, once at about 20-30% colonization and again at about 60-70% for faster growth, if stalled also shake

Bulk substrate:

using straight cococoir or a coco+verm mix works and gives you good results, you can also use dried organic animal manure, coco coir can be soaked and rinsed to wash out any dirt if its not great quality, but in general you want to slowly add filtered or spring water in your substrate and mix it around until when you grab a handful and squeeze it hard you get around 20drops of water MAX, large vermiculite also helps with water retention later on

You should ideally use your sub right away, for smaller grows, you can find a large pot and submerge your bulk substrate bag with a weight underwater (and put something to stop the bag from touching the bottom of the pot) and bring the temp up to 85C and keep it there for 1.5 to 3 hours (longer for manure or ... hotter/nutrient rich material), and take it out to cool somewhere clean before you use it, for larger grows you have to create a large box or barrel pasteurizer which you can find on youtube, its actually not that complicated


fruiting box:

i use a monotub that has a lid which completely stops any air exchange, you can also use various other sorts of boxes, but consider good quality

4 holes each 2.5cm in diameter will be cut, each around the top corners of the monotub, while the spawn and bulk sub are colonizing it should be sealed off with non breathable tape, and when they are fruiting it should be replaced with micropore tape or filters

-wipe the tub Clean with 70% isopropyl and lysol and spread your garbage bag inside and also wipe your bags with 70% isopropyl and lysol and let it dry,spread your black garbage bag in the tub and mix the spawn and bulk substrate 1:2 layer by layer, start with the bulk and also finish with the bulk, don't let any grain be exposed to air on top and finish it to make up a total of 12-13 cm thickness, in front of the flow hood or somewhere with still clean air
close the lid and colonize at 23C-27C

Pinning:

after about 10 to 14 days or where mycelium has fully colonized tubs, slowly reduce the temp to 20c then replace the tapes with micropor, you also need a good amount of white light to hit the surface directly at least a 6 hours a day

Try to maintain temps at 20C-22C which makes for quality fruits

If surface is drying and no small water droplets can be seen spray pre boiled water at the walls of the tub and maintain humidity very high

Harvest:

before veils are opened (not while and not after) is a great time to harvest, concerning aesthetics and potency, fruit bodies can be taken from base of the stem, rotate clockwise and counter clockwise and pluck with ease, cut the bottoms and take them straight to the dryer

Tissue sampling:

-Take the apex fruit (which contains markers you desire) and in a sterile manner bring in the SAB or the hood, cut open with sterile blade (preferably use a tiny blade on a scalpel) and without damaging the tissue gently leave it on the agar dish and sterilize your scalpel again or have 2 ready at all times
taking tissue from different parts of the stem and cap produced different results, i'd like to isolate the most potent section of the mushroom which IME is just where the puff in base of the stem is ending)

-sectoring tissue from cloned agar plates is important, firstly if mycelium is growing puffy or dense , likely pH is not in the correct range, adding more agar (up to 30g/liter) can also improve mycelium integrity and health but is not a main factor of mycelial health
only sector the most healthy tissue from the plates and transfer for master cultures which will be used to expand more mycelium later


Drying:

using heat to dry the mushrooms will be detrimental to their potency and looks, use a dryer without a heat element, within 24h these should be cracker dry, its also important to seal them in vacuum bags with some dehydrating bags straight from the drier to maintain potency and long storage, store in room temp rather than fridge or freezer away from light

Sporework:

-if you have a print, scrape some on the middle of your agar plates (i usually try 6 but you can do more or less with your respective situation) and wait for mycelium to grow, two types of fast and vigorous growth can be spotted as potential fruiters one is a rhizomorphic section and a tomentose section, try to isolate the best tissue on the growth away from the spore drop point to avoid isolating multiple strains

-After isolation on a new agar plate you should see if you isolated a singular tissue based on how it grows and should proceed to spawn both types of mycelium and fruit them in a small box, this is usually to speed up the process of reaching a tissue clone from fruit stem so first we make sure our mycelium fruits and second we don't waste time and resources

-Usually cloning the best fruit from clusters after pinning results in a uniform and strong mycelium, 5 times cloning from fruit is usually the time to take a new spore print just in case the mycelium starts mutating or degenerating, usually when you repeat the process of spore work and fruit tissue work a total of 5-6 cycles your mushrooms look and act significantly different compared to the starting spore batch, this is also the method used to domesticate (whatever that means) wild mushrooms, and mixing different spores on agar and moving on with the process can result in crossing stable genetics, so this concludes general sporework for the home grower

Dedicated to the mushroom and all of its patrons

Espurrr attached the following image(s):
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Live plants. Sustainable, ethically sourced, native American owned.
 
Espurrr
#2 Posted : 9/8/2019 4:57:48 PM




Posts: 403
Joined: 23-Aug-2015
Last visit: 09-Mar-2024
Location: Iran
 
Brennendes Wasser
#3 Posted : 9/8/2019 9:30:00 PM

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Insane Shroomies Shocked Shocked Shocked The last Picture is a true Pinup Cube Laughing Laughing Laughing
 
Espurrr
#4 Posted : 9/9/2019 9:38:00 AM




Posts: 403
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Last visit: 09-Mar-2024
Location: Iran
i've been working on this for 3 years now, this tek is sure to run quick, yield much, and surprise you in terms of potency!
more pictures and videos over time Thumbs up
 
Felnik
#5 Posted : 9/9/2019 5:58:02 PM

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Very scientific I like it
The only way of discovering the limits of the possible is to venture a little way past them into the impossible.
Arthur C. Clarke


http://vimeo.com/32001208
 
Espurrr
#6 Posted : 9/12/2019 3:51:50 PM




Posts: 403
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Felnik wrote:
Very scientific I like it

hi !
about that, im planning to send my agar medium, spawn, Liquid medium, prepared substrate and dry fruit sent for analysis, so when thats done i'll share the specs
what i think would make that significant is I've experimented to predict the most healthy, speedy, high yielding, potent approach to cultivating cubensis, and maybe from the analysis, we can come to a general consensus about the natural preferences of psilocybe cubensis
any other ideas are also appreciated
 
doubledog
#7 Posted : 9/12/2019 4:12:08 PM

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Isnt better to pasteurize casing layer? Instead of autoclaving?
I had big problems with contamination in casing layer when it was autoclaved, once I switched to pasteurization, these issues dissapeared.

Btw, It is quite cool to add some colouring to the agar, it increase aesthetics. I have used beetroot juice for this purpose.
 
Espurrr
#8 Posted : 9/12/2019 4:27:58 PM




Posts: 403
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doubledog wrote:
Isnt better to pasteurize casing layer? Instead of autoclaving?
I had big problems with contamination in casing layer when it was autoclaved, once I switched to pasteurization, these issues dissapeared.

Btw, It is quite cool to add some colouring to the agar, it increase aesthetics. I have used beetroot juice for this purpose.

hi , not sure if the thread is updated for you but
Quote:
pasteurize for 2 hours in 80c and let it drain in a clean environment until 1 or 2 drops fall when squeezed

the humic / fulvic acid i use gives the agar some color, good enough for sighting any contamination
 
doubledog
#9 Posted : 9/12/2019 5:49:40 PM

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I am refering to this section about casing layer
Espurrr wrote:

-Casing layer (optional)
50% vermiculite + 50% coco-coir
add enough water to make damp
autoclave for 6 0minutes at 121C inside a large spawn bag
add 0.5cm to 1cm casing layer while mono-tubs are mostly colonized and create riggid surface with sterile fork (better surface evaporation and air circulation = pins)
 
Espurrr
#10 Posted : 9/12/2019 6:51:38 PM




Posts: 403
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Last visit: 09-Mar-2024
Location: Iran
doubledog wrote:
I am refering to this section about casing layer
Espurrr wrote:

-Casing layer (optional)
50% vermiculite + 50% coco-coir
add enough water to make damp
autoclave for 6 0minutes at 121C inside a large spawn bag
add 0.5cm to 1cm casing layer while mono-tubs are mostly colonized and create riggid surface with sterile fork (better surface evaporation and air circulation = pins)

oh, i haven't had any issues either autoclaved , pasturized, or simply boiled vermiculite chunks drained and spread on the surface
 
doubledog
#11 Posted : 9/12/2019 9:32:18 PM

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Issues which I and my friends have encountered, were always from coco-coir.

However, your tek is definitely great, I have used almost the same approach with excellent results.
 
Espurrr
#12 Posted : 9/13/2019 3:28:12 PM




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doubledog wrote:
Issues which I and my friends have encountered, were always from coco-coir.

However, your tek is definitely great, I have used almost the same approach with excellent results.

makes sense, i wanted to remove coco coir from the tek before but thought maybe its better in a mix
when working with synthetics high grade coco coir seems to do a good job, however
 
Chaska
#13 Posted : 9/29/2019 6:27:14 PM

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amazing. with a new focus on cube cultivation im finding this inspiring. cant wait to see the rest of the pics
grow plants, make tea, love life
 
infinitynlove
#14 Posted : 9/30/2019 6:35:11 PM

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Superb! very accurate!

Couldn't of said better!

There is a tek that allows you to make your agar a lot easier, filter the waste water from boiling your rye grains and use that for your agar, You don't need to add any more nutrients.

But your mix is excellent and your results show in your perfectly formed mushrooms.

Loving it!

<3

I am certifiably insane, as such all posts written by me should be regarded as utter nonsense or attempts to get attention in fact everything I write here is a lie !

I hope in some way, my posts and replies may of helped you, I hope you like what I have said here if not feel free to send me a none flame PM
 
infinitynlove
#15 Posted : 9/30/2019 6:49:28 PM

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Espurrr wrote:
doubledog wrote:
Isnt better to pasteurize casing layer? Instead of autoclaving?
I had big problems with contamination in casing layer when it was autoclaved, once I switched to pasteurization, these issues dissapeared.

Btw, It is quite cool to add some colouring to the agar, it increase aesthetics. I have used beetroot juice for this purpose.

hi , not sure if the thread is updated for you but
Quote:
pasteurize for 2 hours in 80c and let it drain in a clean environment until 1 or 2 drops fall when squeezed

the humic / fulvic acid i use gives the agar some color, good enough for sighting any contamination


Casings, if not done right, are often a source of contamination.

I have had several contams from casing layers when I first got started. Reading the posts of many the trusted cultivators on the shroomery I noticed that they often move away from casing layers, for this reason.

If the sub is really thick, 5" and above, and the humidity is kept above 90% rh, a casing layer often isn't required.

Casing layers are really useful if you have a thin sub, under 3", as they add a layer of moisture that prevents the sub from becoming to dry they also reduce the need for regular misting / spraying.

They are a requirement for pf cakes! the diff in yield can be 100% increase when using dunk and roll + top casing for pf cakes

late casing on bulk often works well, which is simply adding a casing layer once knots are visible.

<3
I am certifiably insane, as such all posts written by me should be regarded as utter nonsense or attempts to get attention in fact everything I write here is a lie !

I hope in some way, my posts and replies may of helped you, I hope you like what I have said here if not feel free to send me a none flame PM
 
Espurrr
#16 Posted : 10/1/2019 3:00:28 AM




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Quote:
There is a tek that allows you to make your agar a lot easier, filter the waste water from boiling your rye grains and use that for your agar, You don't need to add any more nutrients.

hi , yes maybe run off water from millets then added gypsum light malt extract (or even using molasess) , i'd check to see if my agars ph is at about 7 after making it although cubensis feeds in a wide range of ph in mediums but since we're trying to balance the grain and bulk sub on 7 then it'd be better if it always stays in that ph

what i did for mono0tubs this time is in the pictures below
so now i won't let any spawn remain near to the surface of the sub, to get thousands of pins consistently in the monotub
when tubs are 75% i move them to 17C for a day then 23C stable, it stops growing like before and some days later (very soon) you see these pins on the bed
so in my mind im far away from the perfect grow, but someday soon its gonna happen
so i've asked people max dried grams they got from a tub, the answers were mostly ranging between 200-300 g (70L monotub 12.5cm thick beds)
so far 1 time we did something that made this monotub make 300g dry in the first flush , the cake looked sort of destroyed
so when tossing it, there was so many thousand new pins on it and growing, so for whatever reason we tossed those, but some factors were off with that formulation specially in terms of ph and some nutes being way more concentrated than the others, anyway
im thinking if by some tek you can have a monotub that produces 500g dry in 2 flushes, destroying the cake completely, that'd be superb ?
keep getting reminded of somebody writing a paper about growing cubes in a jar in ancient egypt, if they knew whats going on in our apartments now ! Very happy
so anyway, here is cubensis with no casing layer , 7 days from spawn + 4 days in the monotub + 6 days in the fruiting room = 17 days to first pin
Espurrr attached the following image(s):
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20190929_122657.jpg (2,467kb) downloaded 655 time(s).
 
Chaska
#17 Posted : 10/17/2019 12:07:52 AM

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KEEP IT COMING FRENBig grin
exciting to see 86 f incubation temp being so succesful! and fruiting at 76!
grow plants, make tea, love life
 
infinitynlove
#18 Posted : 10/17/2019 2:24:52 PM

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Chaska wrote:
KEEP IT COMING FRENBig grin
exciting to see 86 f incubation temp being so succesful! and fruiting at 76!


I read that 80f is enough for incubation / colonisation as they mycelium generates its own heat and adds approx 5-6f to incubation temp, with 86 f internal temp being ideal.

So an incubation temp of 80f = 85/86f internal cake / jar temp.

I hear higher temps are more suitable to mold, so higher temps = more chance of contams, and no one likes contams!

To end all doubts I should do a side by side where I incubate some mold at diff temps and incubate some shrooms at the same temps to find the optimum growth of mycelium vs mold growth.

80f has worked great for me so far, whatever works for you.... fruiting at 76f is ideal, but once fruiting you can raise the temp for faster shroom growth, but it makes weaker shrooms. ime slower shroom growth = stronger shrooms.

Oh and don't let them sit in the sun when growing, ime it reduces potency considerably.

Peace <3


I am certifiably insane, as such all posts written by me should be regarded as utter nonsense or attempts to get attention in fact everything I write here is a lie !

I hope in some way, my posts and replies may of helped you, I hope you like what I have said here if not feel free to send me a none flame PM
 
Espurrr
#19 Posted : 10/19/2019 2:24:08 PM




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81F is a good temp to inoculate, i work in a designated room for mushroom work so everything is clean and in 86F i have 1% or less contamination rates
you could argue that 86F will cause genetic deterioration, this may be true ( and i'll change the tek if i come to that conclusion or someone illuminates the information)
slower formation of pinset and fruit body growth (10 days instead of 5) is key ime, which is why i shock at 17C and keep at 23C
 
infinitynlove
#20 Posted : 10/21/2019 7:54:51 AM

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Espurrr wrote:
81F is a good temp to inoculate, i work in a designated room for mushroom work so everything is clean and in 86F i have 1% or less contamination rates
you could argue that 86F will cause genetic deterioration, this may be true ( and i'll change the tek if i come to that conclusion or someone illuminates the information)
slower formation of pinset and fruit body growth (10 days instead of 5) is key ime, which is why i shock at 17C and keep at 23C


I would be interested to know more about genetic deterioration, I have noticed old spores and old mycelium show signs of senility, which is down to genetic deterioration over time, but I was not aware that this would happen with a slightly higher temp of 86f ?

Agreed Lower fruiting temps are a must for high potency and high density mushrooms.

Loving the tek and the shroom picks Smile

inf <3
I am certifiably insane, as such all posts written by me should be regarded as utter nonsense or attempts to get attention in fact everything I write here is a lie !

I hope in some way, my posts and replies may of helped you, I hope you like what I have said here if not feel free to send me a none flame PM
 
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