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Ullr
Stranger

Registered: 10/27/17
Posts: 93
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First P. cubensis cultivation log - shoebox #3 harvested, #4 and #5 flushing (Z strain) 1
#24746121 - 10/29/17 06:16 PM (6 years, 4 months ago) |
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I intend to grow cubes for the first time. I plan to start with an agar culture from a spore syringe, clean up possible contams, inoculate to rye and spawn to a 45-liter monotub. I may directly inoculate some rye jars if there's anything left in the syringe after agar and try to get something growing while I clean and isolate cultures.
Unfortunately, things got delayed and cold weather is nocking the door. Indoor temperatures are currently ranging 13 to 18 Celsius, 15-17 most of the day. That's about 55 to 65 F. I'm living in an apartment without heating, and can't really afford to heat up the house given how terribly insulated it is, so that's not an option.
I've searched around and read local heating is not a good idea, but will cubes grow at all at these temperatures, that will most likely have dropped further to 50-60 F when fruiting time comes? Between cold and local heating, which would be the lesser evil?
I'm considering cultivating in a heated big closet, by insulating with styrofoam opening small vent holes at the lower and upper ends of the closet, placing the tub at a rack in the middle/top, and a big heating mat at the bottom, and letting the warm air current provide some FAE. Like a monotub inside a heated monotub. Would that be a viable option?
Thanks in advance! _____________________________________________________________________________
UPDATE:
I gathered all the supplies and ended up using a smaller and crappy closet for the first experiment. The door has quite a big gap, particularly at the bottom part, and it already had some holes drilled in the top. The only modification was a new hole for the cable and a new shelf full of holes.
I got ahold of a small thermostat controlled oil heater, and place a LED light with a timer set 12/12 (LED light has quite a wide non-visible spectrum so it should be pretty close to natural for this purpose). It seems like it keeps the temperature just fine (only turning on for a short time every once in a while), plus it keeps the lower part about 5 Celsius warmer than the upper, so I can have jars colonizing in the lower shelf and a tub in the upper one at the same time.
Only the syringe (Z strain) is yet to arrive. I expect to be inoculating agar plates and some jars in a few days.
Any thoughts about the setup?
 _____________________________________________________________________________
11/15/2017
Well, I was delayed by some issues but I finally got to prepare everything. I'm using rye in 18 tall and slim 1/2 pint jars, since my PC is pretty narrow and I found I could get a bigger amount of spawn per PC run this way, plus I already had the jars. Using polyfill as a lid filter. One of the jars cracked for some reason, while another got a lot of burst grains at the bottom. The rest are looking fine, with fat, dry-looking rye.
Agar is being sterilized right now, and the SAB is ready with some glass petri dishes inside awaiting pouring.
Photos soon. _____________________________________________________________________________
11/17/2017 The promised photos. There are around 3 1/2 quarts of inoculated spawn in the 18 jars:

Here you can see the cracked jar. I've left it to see what happens, since the crack is not open and there's no way gas is going through it. It may even stay clean.

The jar that had a bunch of grains burst at the bottom during PCing:

The rest are all looking like this:

Some of the petri dishes got bad condensation after inoculation, I'm guessing temperature drop. It did fix somewhat after the temperature inside the closet stabilized. I hope it doesn't prevent growth, but even if it all fails I still have enough in the syringe for quite a bunch of petris.

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11/24/2017
I'm getting some bacterial nasties in my agar plates, presumably making their way through the polyethylene film via ridiculous amounts of condensed water.
A couple of days ago I noticed the tiny bacterial dots at the edges of my plates and prepared everything for transfer but, unfortunately, I had some unexpected business to attend and had to drive off after pouring some new plates.
I did the transfer today and contamination had extended a great deal. Also, my mycelium looks weird, way too fluffy, may that be because of the excess humidity or lack of gas exchange? Polyethylene is supposed to allow gas transfer, but with all the condensation most plates were closed shut, lid stuck to plate by vacuum.
An example of the condensation on a plate where no mycelium grew:

The contaminated plates:

Closeup on the mycelium (I'm guessing it's mycelium, I hope I'm not transferring some weird crap):

The grain jars show no sign of growth or contamination so far.
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11/30/2017
I'm now in the 3rd transfer and a couple of transfers are showing some nice, aggressive rhizo growth.
The second transfer was a total mess. I used a 20 minute PC cycle for both the agar and petri dishes. It must have not worked, because the following day I had like three kinds of bacteria everywhere, inside, over and under the agar.
On the third transfer, two plates have been contaminated so far, both via the scalpel and only where the transfer was placed, meaning I picked some sneaky bacteria along with the mycelium.
Jars are showing the first signs of growth, let's see how that goes. Photos tomorrow. _____________________________________________________________________________ 12/01/2017
3rd transfer plates starting to show some nice rhizomorphic growth:

Not-so-nice 3rd transfer plates where my scalpel blade picked some hidden bacteria:

Another scalpel contam, this time from the 2nd transfer. This started off as a huge line of bacteria with a tiny mycelium spot in the middle. I can't believe how agressively this mycelium has fought the contaminant, it grew straight over it with rhizomorphic growth and only once it had the bacteria in control it started to produce the thinner and wider growth that can be seen now. It's kinda fun to see how these little organisms interact and compete:
 _____________________________________________________________________________ 12/11/2017
Current stage of rye colonization:
 Over a month after inoculation and only 7 out of 18 jars are showing growth, most of them starting last week, none new in the last 4 days.
I'm now in the 4th agar transfer, ready for the 5th. Which plate/sector would you say looks best?
These are the best 3rd transfer plates... I'm leaving these to see if I can get some pins to clone:

_____________________________________________________________________________ 01/05/2018
I wasn't able to post in a long while, this is been a busy month and I didn't even get the time to take care of the cultures as desired, least to take photos and update this.
The 5th transfer was made to tea agar soon after my previous update. I still had a couple of contaminants growing from the wedges, but they didn't spread. Most of the jars didn't ever get any kind of growth from the MS inoculation, neither did most of the initial petri dishes. It looks like my syringe didn't have many viable spores.
Thus along with the transfer, I took wedges from some discarded (slower, fuzzier) yet clean plates and threw them into the jars that weren't growing anything.
The good part of a syringe with few viable spores is that I may already have an isolate or something close enough for my goals. I didn't take a photo before the mycelium spread all over the place, but this plate looked really even.

Not so even, nor faster, but pretty thick rhizo growth on another 5th transfer plate:

On the 2nd January, I finally got some time to finish up business. I started by changing my led to a 6000k bulb. Since three 100% colonized jars had been already sitting there for two weeks while the rest were between 60 and 80% (grains drying out = slow colonization), I decided to dunk and spawn those three to a shoebox with CVG. All seemed and smelled clean:

This is the shoebox 9 hours after spawning:

About 40 hours after spawning:

The remaining jars. These will be spawned to a monotub, unless contaminated:

One of them shows suspicious activity (it being horribly dry aside). It's kinda weird, would you say this is contamination or the metabolites may have other reasons to appear?:

I also found that moving my petri dishes to the lower part of the closet reduced temperature shifts and thus completely eliminated my condensation problem, while keeping them a bit colder yet pretty close to room temp.

Also, my goal-culture spores arrived:
 I've already inoculated several plates with these. I've also done the 6th cubensis transfers today, which unless problems arise will become my set of master plates (for now).
Happy new year!
_____________________________________________________________________________ 01/10/2018
This thing is knotting like crazy, although the mycelium looks a bit uneven in thickness. Should I put it in fruiting conditions?

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This is going fast!

_____________________________________________________________________________ 01/11/2018
Pinning:

_____________________________________________________________________________ 01/12/2018
Pinset update. More primordia popping out as well. Is it me or it looks a bit too dry?

_____________________________________________________________________________ 01/13/2018
Fast indeed. The biggest ones here are 2 inches tall, but I'm noticing a couple of issues. Namely the fuzzy feet, and the fact that the veils are tearing already on some, while the caps look quite underdeveloped. I did a search and couldn't come up with a clear answer, some say it may be FAE, some say humidity, some say genetics. Any ideas on what I can do now to prevent more veils from opening prematurely, or what to do better for the next flush?

_____________________________________________________________________________ 01/14/2018
Today I picked up my first shrooms. Tiny, but better than nothing I guess. The rest of open caps will be picked tonight, I wouldn't want to wake up to a mess of spores.

I've also spawned a second shoebox from three fully colonized jars. No liner this time, just to see how it goes. The remaining jars are pretty even, I'll spawn those to a full-sized monotub.

_____________________________________________________________________________ 01/15/2018
Harvested the second stage of the first flush. There are still some slower ones that should be ready tomorrow.

The "big" ones:

The tiny ones:

_____________________________________________________________________________ 01/18/2018
The first shoebox started smelling somewhat moldy, so I've put it aside just in case. No visible mold, and I'm not noticing the smell today, so maybe it will still give a second flush.
The second shoebox is colonizing just fine:

And I decided to spawn the remaining jars to more shoeboxes:

_____________________________________________________________________________ 01/22/2018
The second shoebox (middle) is knotting now, while two of the later ones are almost there. The left one I packed down a bit, and it seems to be getting a more even colonization (as seen from the sides and bottom). Experimenting a bit.

This smaller one was spawned from three jars that never made it to 100%, and stalled at maybe 95%. The top grains were rock dry. I shook those uncolonized grains loose, tossed them, and dunked and spawned the rest separated, just in case some uncolonized grains made it to the sub or contaminants were present.
There's aereal mycelium and slower surface colonization, but the top layer was thickish so not sure if it may be bacterial or not.

_____________________________________________________________________________ 01/25/2018
The first shoebox got the green + some fuzzy gray mold and got tossed. Glad I had taken it out of the grow area.
Now three out of the remaining 4 shoeboxes are showing hyphal knots. I should have some more fruits for the next week.
_____________________________________________________________________________ 01/30/2018
Now this leaves me baffled. Two of the shoeboxes are pinning. One spawned the 14th January and the other the 18th. Both have been kept in the same closet, same temperature, same water content, same spawn ratio, same fruiting conditions (reversed lid if I'm not around, since pinning open air and mist as the beads dry if I'm around to take care of it). No signs of contamination that I'm aware of, no metabolites, no bruising, no outreaching mycelium and smells good.
Both have some side pins, not many, there's not really a gap big enough for them to grow. But the later one has hundreds of top pins and primordia (including a few blobs/mutants), while the earlier one has 15 or so pins, and a big one that just went from pin to opening the cap with no veil developing.
As for the differences, the earlier one has almost no top layer (forgot to set aside some CVG before mixing the spawn), I leveled the sub without packing it down, as usually advised.
The later one has a thin top layer and was lightly packed down.
Earlier shoebox with its single shroom:

Later shoebox. Nevermind the crater. Someone accidentally pushed me while taking photos and I shoved my phone into the sub 

So... help me here, please. Did I do something wrong that can explain this? How are my surface conditions looking like? Crappy genetics or environmental factors?
_____________________________________________________________________________ 02/01/2018
Pinset update for the 2nd shoebox. Not really a flush, just some pins randomly popping here and there every once in a while. The caps opened even earlier on these (which has been discussed here), and I already had to pick three dwarf shrooms, plus it's getting plenty of side pins now.


Pinset on the 3rd shoebox is looking far better despite the phone incident, with some interesting pale genetics. It has a single, tiny side pin. I'm seriously puzzled about the difference.

_____________________________________________________________________________ 02/05/2018
Finally got some spare time to update this.
Shoebox #2 was tossed after its few pins aborted and it started smelling funky. Dry yield: 0,5 gram  No big deal though. Let's not forget these are MS to grains, and probably bacterial to whatever extent.
Shoebox #3 was harvested a couple of days ago. The fruits were thicker and denser than those on box #1, plus light pigmented but, unfortunately, most pins aborted. I took a couple of clones from the bigger fruits. There was one single side pin. Dry yield: 12 grams, 3 of which were aborts.

Also got some mutants:

Shoebox #4 is pinning really unevenly. First pin appeared days ago, later on a couple more, and today I found 5 or 6 popping up from the surface. No clustering whatsoever, just random isolated pins and a ton of fat side pins. That first pin is already the biggest shroom I've grown to date, and is showing some interesting PE-like traits. I'm cloning it:

Shoebox #5 is also starting to flush, throwing some slim shrooms. Pinset is still filling in:
...and it's side and bottom pin city:

It looks like this "Z strain" syringe came with an interesting variety of genetic traits.
I also inoculated some grains with the clones via agar.
Edited by Ullr (02/05/18 09:45 AM)
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Morel Guy
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Registered: 01/23/13
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Re: Cold weather P. cubensis cultivation [Re: Ullr]
#24746175 - 10/29/17 06:33 PM (6 years, 4 months ago) |
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The old way was a water bath tub set up with temp controlled aqarium heater. Would recommend trays or cakes for fruiting. I also had a cold room.
-------------------- "in sterquiliniis invenitur in stercore invenitur" In filth it will be found in dung it will be found
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mushboy
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Registered: 04/24/05
Posts: 32,502
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Re: Cold weather P. cubensis cultivation [Re: Ullr] 1
#24746177 - 10/29/17 06:34 PM (6 years, 4 months ago) |
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cold can slow down growth but its not impossible. 70f would be better.
these fruited in 60f
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Citric



Registered: 03/19/04
Posts: 4,490
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Re: Cold weather P. cubensis cultivation [Re: Ullr]
#24746178 - 10/29/17 06:34 PM (6 years, 4 months ago) |
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Will they grow? Sure. But slowly.
You can get a tiny space heater for rather cheap at Wal-Mart or stores alike.
-------------------- Self Healing lid tek ** Update 10.17.17 ** Mini casing pictures: Pins to harvest Cup O' Shrooms Magash: I noticed my contams were in the shape of fingers Hyphae: Yes "Loss of moisture from the substrate" is not a casing trigger. My final Grow!
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Morel Guy
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Re: Cold weather P. cubensis cultivation [Re: Citric]
#24746188 - 10/29/17 06:37 PM (6 years, 4 months ago) |
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Not good for spawning and I read fruits will be slower and larger.
-------------------- "in sterquiliniis invenitur in stercore invenitur" In filth it will be found in dung it will be found
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hamloaf
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Registered: 12/23/09
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Re: Cold weather P. cubensis cultivation [Re: Morel Guy]
#24746189 - 10/29/17 06:38 PM (6 years, 4 months ago) |
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Higher temps produce larger fruits. Colder temps produce short, dense fruits.
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Ullr
Stranger

Registered: 10/27/17
Posts: 93
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Re: Cold weather P. cubensis cultivation [Re: mushboy]
#24746201 - 10/29/17 06:46 PM (6 years, 4 months ago) |
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I'd colonize in the same heated closed, and place a light I still have from a reptile terrarium on top, BTW.
Quote:
mushboy said: cold can slow down growth but its not impossible. 70f would be better.
these fruited in 60f

Those are beautiful! Which variety is that?
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mushboy
modboy



Registered: 04/24/05
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Re: Cold weather P. cubensis cultivation [Re: Ullr] 1
#24746204 - 10/29/17 06:47 PM (6 years, 4 months ago) |
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dont remember.
genericious cubensis
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Ullr
Stranger

Registered: 10/27/17
Posts: 93
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Re: Cold weather P. cubensis cultivation [Re: Morel Guy]
#24746208 - 10/29/17 06:50 PM (6 years, 4 months ago) |
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Quote:
Morel Guy said: Not good for spawning and I read fruits will be slower and larger.
So I assume they will fruit, just slower. But will it prevent colonization/spawning? I'll be using coir+verm.
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Morel Guy
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Re: Cold weather P. cubensis cultivation [Re: Ullr]
#24746284 - 10/29/17 07:37 PM (6 years, 4 months ago) |
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That is cold for spawning.
Lot's of items can be less than ideal and give a return.
-------------------- "in sterquiliniis invenitur in stercore invenitur" In filth it will be found in dung it will be found
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Apples in Mono
Not a puppet


Registered: 09/21/17
Posts: 3,240
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Re: Cold weather P. cubensis cultivation [Re: Morel Guy]
#24746304 - 10/29/17 07:46 PM (6 years, 4 months ago) |
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There will be plenty left in the syringe after putting a few drops to agar; you'll still have almost all of it
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Gurdjieff
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Registered: 10/02/17
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Re: Cold weather P. cubensis cultivation [Re: Apples in Mono]
#24746341 - 10/29/17 08:02 PM (6 years, 4 months ago) |
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You dont have a closet or bathroom you can heat and convert into your fruiting lab?
-------------------- Man lives in sleep.
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Ullr
Stranger

Registered: 10/27/17
Posts: 93
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Re: Cold weather P. cubensis cultivation [Re: Gurdjieff]
#24746388 - 10/29/17 08:22 PM (6 years, 4 months ago) |
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Quote:
Gurdjieff said: You dont have a closet or bathroom you can heat and convert into your fruiting lab?
Sure, I quote myself:
Quote:
Ullr said:I'm considering cultivating in a heated big closet, by insulating with styrofoam opening small vent holes at the lower and upper ends of the closet, placing the tub at a rack in the middle/top, and a big heating mat at the bottom, and letting the warm air current provide some FAE. Like a monotub inside a heated monotub. Would that be a viable option?
What I lack is insight on the viability of this setup, or a guideline to follow.
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Gurdjieff
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Re: Cold weather P. cubensis cultivation [Re: Ullr]
#24746980 - 10/30/17 07:05 AM (6 years, 4 months ago) |
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Sorry I read it when I was half asleep. Get a thermometer and a heater and test the closet, see how well it retains the heat you need and how long. You might not need to insulate if it holds the temps you need.
-------------------- Man lives in sleep.
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Ullr
Stranger

Registered: 10/27/17
Posts: 93
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Re: Cold weather P. cubensis cultivation [Re: Gurdjieff]
#24748449 - 10/30/17 07:47 PM (6 years, 3 months ago) |
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Quote:
Gurdjieff said: Sorry I read it when I was half asleep. Get a thermometer and a heater and test the closet, see how well it retains the heat you need and how long. You might not need to insulate if it holds the temps you need.
Thanks for the tip!
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Yonatin


Registered: 09/05/17
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Loc:
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Re: Cold weather P. cubensis cultivation [Re: Ullr]
#24748616 - 10/30/17 08:56 PM (6 years, 3 months ago) |
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I currently fruit in 63-70 degree temps just fine. They aren't super tall or anything but like everyone else has said they are dense. one of them weighed 2.3g just by itself. And for colonizing I use a TIT to keep the temps around 76 degreees and haven't had a problem so far.
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Ullr
Stranger

Registered: 10/27/17
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Re: Cold weather P. cubensis cultivation [Re: Yonatin]
#24748648 - 10/30/17 09:11 PM (6 years, 3 months ago) |
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Quote:
Yonatin said: I currently fruit in 63-70 degree temps just fine. They aren't super tall or anything but like everyone else has said they are dense. one of them weighed 2.3g just by itself. And for colonizing I use a TIT to keep the temps around 76 degreees and haven't had a problem so far.
I'm at 55-65 and it will most likely have dropped to 50-60 by the time it fruits. I was actually planning to heat just enough to prevent temperatures from falling under 60.
Also, what's a TIT? I know, search engine, but I have limited internet data available this week.
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pixelpopper
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Re: Cold weather P. cubensis cultivation [Re: Ullr]
#24748659 - 10/30/17 09:17 PM (6 years, 3 months ago) |
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tub in tub
Quote:
TiT - "Tub in Tub", refers to an incubator consisting of 2 plastic tubs and an aquarium heater.
Edited by pixelpopper (10/30/17 09:19 PM)
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Yonatin


Registered: 09/05/17
Posts: 654
Loc:
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Re: Cold weather P. cubensis cultivation [Re: Ullr]
#24748683 - 10/30/17 09:29 PM (6 years, 3 months ago) |
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Quote:
I'm at 55-65 and it will most likely have dropped to 50-60 by the time it fruits. I was actually planning to heat just enough to prevent temperatures from falling under 60.
Also, what's a TIT? I know, search engine, but I have limited internet data available this week.
Yeah like Pixel said it's two totes or tubs whatever you want to call them. You fill one up with water and place an aquarium heater in it (a fully submersible one) and then place the other tote in the one full of water. Then the water heats the bottom which heats the air in the tote. If you can get temps in the 60's you should be fine they will just grow slower.
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Ullr
Stranger

Registered: 10/27/17
Posts: 93
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Re: Cold weather P. cubensis cultivation [Re: Yonatin]
#24755275 - 11/02/17 06:45 PM (6 years, 3 months ago) |
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I gathered all the supplies and ended up using a smaller and crappy closet for the first experiment. The door has quite a big gap, particularly at the bottom part, and it already had some holes drilled in the top. The only modification was a new hole for the cable and a new shelf full of holes.
I got ahold of a small thermostat controlled oil heater, and place a LED light with a timer set 12/12 (LED light has quite a wide non-visible spectrum so it should be pretty close to natural for this purpose). It seems like it keeps the temperature just fine (only turning on for a short time every once in a while), plus it keeps the lower part about 5 Celsius warmer than the upper, so I can have jars colonizing in the lower shelf and a tub in the upper one at the same time.
Only the syringe is yet to arrive. I expect to be inoculating agar plates and some jars in a few days.
Any thoughts about the setup?

Edited by Ullr (11/02/17 06:47 PM)
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