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Offlineshroomator
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Workflow?
    #26097777 - 07/09/19 09:21 AM (5 years, 6 months ago)

Ok I am about to start my first monotub. :smile:

Currently wondering what the workflow will be afterwards. Do you soak the monotub after each flush?
How many flushes would you recommend?
When can I start getting more grain ready?
Would be nice to start directly when the jars are empty but I guess then they are ready and I have no empty monotub.

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Offlinejbgtaa
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Re: Workflow? [Re: shroomator]
    #26097790 - 07/09/19 09:35 AM (5 years, 6 months ago)

I just finished a mono tub as well, and been thinking alot about propagation and just basically keeping shit rolling. Heres where i went from my mono harvest, keep in mind before this route im working on, i was going from spores to grain to mono, and as a beginner that may seem like the fastest way (it did for me), but really its not. Keeping a solid culture going speeds things up immensely. Here we go:
1. Find your best fruit from the 1st flush and clone it. If you absolutely wont work with agar, then you can use the print, but that almost defeats the purpose of what im about to lay out for u.
2. From a different solid fruit, take a spore print as either a last resort back up or to make a syringe for agar, or a spore syringe for direct MS inoculation. Definitely can and most likely will give off fruits, but theres no way to accurately predict appearance, quality, or the weight of what it puts out. You're going blind with regards to appearance, growth characteristics, and in the very worst case, potency.
3. After your clone genetics are satisfactory (TO YOU. You decide what you want in fruit/agar characteristics), Inoculate master grain jars.
      3A. If you really are digging your plate, go ahead and slant that for use as a back-up master culture. Some may say this is too early, but it's young growth and thats always better as a master than something that has been through fruit multiple times.)
4. Inoculate Master grain. Go LI, LC, GLC, Agar drop, whatever you feel comfortable with not contaminating.
5. When these master jars are near 100%, go ahead and make some recieving jars. Theres alot of teks on G2G if you search. Just make sure u have clean spawn.
6. Fruit your receiving jars in a mono (if thats what you prefer, but anecdotally proven to be the best way to fruit grain).
Thats it. Optional 7th step would be to make your receiving jars +1, and use that +1 jar as another master to keep propagating your spawn. Just remember to throw in some isolated MS to switch it up a little bit every now and then. This is real hobbyist shit, and if you do this once you will have enough mushrooms to last you for a very long while, meaning that you dont have to keep this shit up if your satisfied with your first yield. I'm planning on following this route for a while to keep shit really rolling, but its not neccesary once you become satisfied with your stash.


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Edited by jbgtaa (07/09/19 09:39 AM)

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Offlinesyroth
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Re: Workflow? [Re: jbgtaa]
    #26097960 - 07/09/19 11:26 AM (5 years, 6 months ago)

First off this account is new but I am not new to this process by any means. Secondly, I wanna specifically mention that I agree with jbgtaa's advice.

    I wish to contribute the idea that each process must be tailored to your preferences and your goals/expectations. The aggressiveness of the timeline such as how quick to begin the next phase of something ( new grains after you empty your grain jars into a tub ) is entirely your preference. Aside from avoiding contamination... most of the choices we make in the process are largely a matter of preference based upon personal pet peeves as well as the expectations you set for yourself. In other words, no one can possibly provide you a direct answer without knowing more info such as your goals expectations regarding personal time required and desired output.

    Beyond that let me add some actual specifics for you. There are 2 approaches regarding re-use of existing 'genetics'...

    **One choice is to just use multi spore every time, and not bother with any isolation or cloning techniques. So choosing not to re-use specific genetics but to start 'new' aka multispore/un-isolated/etc genetics.

    **Second choice is to select genetics that appeal to you, then create a plan which balances how long / how much you wish to use those selected genetics compared to the effort taken to do so.


    Let me directly answer what you did specifically ask..

1) I soak tubs only when I feel they are too dry after first flush(es) BUT are still perfectly 'fresh'. Soaking a tub with any contaminants will spread the contamination. Rough weight 'by hand' is your best bet... yes that means you must get a feel. It takes time to get a hang of it because of the time period between spawning to tub and harvesting said tub, you will just have to personally experience it.

2) Depends on sub type / size, mushroom type / size, etc etc. I typically dont bother trying after 3 or 4 flushes. Up until then I will let a tub keep going until I noticed the fresh mushroom smell has left or ' myco piss ' ( sign of infection ).

3) More grains depends solely upon what materials you have and your desired goals. If you are able to start more grains then certainly feel free to do so. Rule of thumb, each phase can easily take a month depending on conditions. Can be way faster of course, but lets say conditions are not ideal such as lower temps for example, then each single step can take weeks. This is my personal rule of thumb when I plan out stages / phases. It aires on the side of caution.


                                                ++++++ Misc thoughts ++++++

    The way I grow I prefer to keep backups of genetics at each stage, and store them in a refrigerator. This means I cold store Master Slants, Master Plates, Master Grain jars, hell even master grain bags and master clones. Please do not forget that 'old age' happens from repeated cellular reproduction. In other words the ULTIMATE GOAL when trying to preserve any set of genetics is to go from SPORE to LONG TERM COLD STORAGE in as FEW steps (transfers) as humanly possible.

    I typically pick some genetics to preserve ( the how / why is irrelevant for this discussion ) and then clean them up on agar. Then I pour a couple of THICK nutritious agar plates. I transfer the genetics to these 'master plates' and then I tightly seal them with parafilm and put them in a ziploc in the fridge. I use 'master plates' because I find, in practice, they last plenty long enough for my purposes. Master slants last longer than master plates.

    Then I, from the same source I used to make the master plates, create a few jars of 600-800 ml liquid cultures. I grow each jar in room temp for a week or two then test on a few agar plates. If it tests clean then I put all but one jar into the fridge for long term storage - (fridge temps slow metabolism down makes them live longer than room temp).

    The jar I do not refrigerate is now my working jar. I then pressure cook agar simultaneously with syringes ( fully wrapped ). I'm skipping details of sterile procedure here but ultimately once agar is poured and cooled I then use 1 syringe to withdraw 5 ml of liquid culture which is wayyyy more than enough for 25 agar plates ( my standard batch ). Each plate needs only ONE drop... although true confession here I will often use 2-3 drops, without any great logical basis for doing so.

    Sometimes if I'm feeling quite confident in a culture (and my sterile technique) then I will go from LC straight to grains. Sometimes I will store many of my first generation of agar plates in room temperature long-term, using them to start new master grain bags every little while. Sometimes not. On that note, sometimes I Grain to Grain religiously... sometimes I skip it entirely. This paragraph is full of preferential equivalents... each has pro's and con's but are largely used or decided upon based on your personal PREFERENCE.

    The fastest cycle I've found involves using a GLC ( grain liquid culture ) straight into grain bags in front of a flow hood. So Spore --> Master Agar ( slant or dish ) --> Master GLC --> Spawn Bags. Best case I harvest in less than 30 days from spore inoculation. Worst case you sneeze at the wrong time and ruin everything :smile: ahahaha

    In your mind, try to work on categorizing different ways of growing so you can learn which steps are interchangeable.

    For example... You can skip agar entirely if you wanted. If so, then above when I refer to 'agar' just sub that for 'master grain jar'. You can cold-store Master Grain Jars just the same as agar plates or slants or liquid culture.

    A "master" ANYTHING, is simply the culture you have at whatever stage / in whatever form... which has been transferred / grown / expanded / cloned the least and you are now using as a source of culture. Could be a master agar plate, master grain jar, master slant, etc etc. You could even have a master PF Cake if you really wanted to!

    None the less, your end goal is always to create 'master' supplies of clean genetics that you can store cold and repeatedly take samples/cultures from over time. Final warning, do not forget I did describe using 'advanced techniques' in this post... for example I do not recommend liquid cultures until you no longer feel the need to ask for help ( in other words until you have the hang of growing without liquid cultures .

    If you have follow up questions, please feel free. But also include basic info such as which is more important working with your existing resources or streamlining your grow? In other words can / will you make a purchase, or should we make a plan based upon what you already have? Also state if you intend to use agar or not, and if you plan to use a flow hood or a SAB? These are the biggest factors we gotta establish prior to diving deeper.

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Offlineshroomator
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Re: Workflow? [Re: syroth]
    #26099716 - 07/10/19 06:39 AM (5 years, 6 months ago)

Thank you two very much for your answers. Exactly the input i needed :smile:.
I started from a spore print --> syringe --> to jar. The spore print i made was really bad but so far it seems like everything ist working so i planned to continue this route.

However working with this stuff really is fun so i want to do it the right way and starting with agar.
I`ve thought the main advantage is just a lower contamination rate, so i didnt really considered it till now. However it seems you also get a better yield and more possibilites to work with it time wise.

How long can i store an agar platae with mycellium?

I have a SAB, its only 40 quart but i managed a grain to grain already. So it should work for pouring some agar dishes.

What does LI, LC, GLC stand for?
I assume LC is liquid culture.

First i want to start with agar but i am going to read deeper in it. Really interesting stuff :smile:. I just like to streamline processes and doing things the proper way. I assume stash should be fine after my first harvest but i am just going to experiment with growing.

Last question:
i got around 12 quart of rye spawn and 2x 40 quarts mono.
How much coir/spawn would you add?

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OfflineSoccrates
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Re: Workflow? [Re: shroomator]
    #26099824 - 07/10/19 09:25 AM (5 years, 6 months ago)

12-24


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Offlinesyroth
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Re: Workflow? [Re: shroomator]
    #26100011 - 07/10/19 10:34 AM (5 years, 6 months ago)

Hey! It should be assumed that one must follow 'sterile procedure' meticulously at all times. In all my answers I wont specifically state sterile technique as I expect readers to assume it is always used.

                                          Agar stuff.....

    Best / Longest lasting would be a thick AND extra-nutritious agar plate which, once inoculated, is promptly wrapped with parafilm and then once colonized stored at fridge temps. This will stay viable up to 18-24 months with confidence... Although as with all things in life nothing is 100% and your mileage may vary. Thicker poured plate plus extra nutrition helps fuel and moisturize the mycellium living on said plate, over the long term.

    Other end of the agar plate spectrum.. you could have a unwrapped super thin low-nutrition agar plate which would last on the shelf a few months at MOST. Again, your mileage may vary of course... if room humidity is 20% versus 80% then you will see a huge difference. I typically see thin plates die from dehydration first. Many times you can re-hydrate the mycellium.

    Agar plates (all plates) maintain cleanliness because of the fact that the vast majority of contaminants fall or waft DOWNWARD onto 'things' - in this case an agar dish. So the extremely tight fit between the top and bottom sections keeps germs from falling in.

    Wrapping in parafilm helps with preventing accidental opening of the plate, as well as in retaining moisture, while simultaneously letting gas exchange. You can asphyxiate your mycellium easily if you were to, lets say, wrap a plate in scotch tape or something of that nature.

    On plate nutrition levels. Less nutrition and a thinner poured plate means more visibility and typically more rapid growth. This is ideal for selecting genetics as well as isolating away from contaminants. Inversely, a thicker more nutritious plate is ideal for long term storage of proven genetics (clean) because we care less about visibility of the mycellium and more about providing food and nutrition for months to come.

    Side note: A professionally prepared Master Slant can purportedly last years and remain perfectly viable. I cannot personally attest to this, but there are experts far more intelligent than myself who have spoken on such matters both here on shroomery but also out and about on the interwebs!

   
                                Onto your other answers.....

    LI = Liquid Inoculant
    LC = Liquid Culture (as you suspected)
    GLC = Grain Liquid Culture

    Many of these terms can be mixed up. For example you could have a MGLC, or master grain liquid culture. But once you suck some into a syringe with intent of INOCULATING something, you have then created a syringe of Liquid Inoculant. So LI is part description of material but also part description of that materials intended usage.

    I typically use 5-7 quarts of spawn per tub. I typically use 325 grams dry coir per tub. Actually thats misleading... I typically like to use a higher amount of spawn in comparison to CVG (coir vermiculite gypsum).

    A tubs yield is most dependent upon its nutrition... which is almost SOLELY provided by the Grains themselves. The CVG substrate is actually fairly low in nutritional content, in comparison to the spawn medium, which is essential in triggering the mycellium to produce fruit.

    Due to this, you can use less CVG material per tub, resulting in a thinner layer in the tub itself. Right now what I ACTUALLY typically do is use 2.5 FULL to brim quart jars of DRY (never measured wet) Rye Grass Seed per tub as a minimum. With this I add ~250 grams (aka slightly less than half of one standard coir brick which is ~650 grams) of dry coir. I used dry and wet just now in potentially confusing ways, i measure all these materials in their dry form prior to usage as a general FYI. Oh, and I use grow bags not quart jars.. so keep that in mind if some of my explanations don't make sense at first.

                      In response to some of your general commentary...

    On agar, you wanna start by absorbing all material on PastyWhites "Pasty Plates", and also be sure to read the "Tiger Drop" sub-section. I also gotta recommend Frank Horrigan and Transcending Life, in addition to PastyWhite. I've found these three to have created huge amounts of accurate material in such a way as to make it easily digestible for the masses. Of course there are so many more I could mention, but I wanted to refer to a few great places to start at first.

    In general, agar does have lower contam rates, as well as a much higher degree of confidence. It also allows you to create as much redundancy in form of genetic 'back up copies' as you desire.

    In many cases, Multi-Spore grows can yield A-LOT! Sometimes they can suck. Its up to you to decide if, over time, you want to see if you can "find a good isolate" which will consistently yield better in some manner (faster, more, etc). Isolates can and often DO produce less. This is because until you TEST what you have isolated you cannot really know what you are isolating. So much more nuance here, forgive my brevity, but I'm leaving Isolating at this level of detail for right now.

    For that just stated reason, many prefer to clone from a particular fruit body aka mushroom. Again, this whole process requires some specific study on your part just in terms of sterile technique ALONE, let alone the cleaning of the culture and subsequent isolation that can follow. However, the idea is you can pick a healthy vigorous and large mushroom and clone that as a starting point in your quest for a 'better isolate'.

    OK! Think I've finished this reply at this point. Again, such a deep complex topic we are only touching the surface of. Feel free to keep asking questions, for brevity's sake I did oversimplify some sections so keep that in mind as you learn more on your own!

    Good luck,



****EDIT: Afterthought. PastyPlates are very SAB friendly as you seal them individually with LIQUID agar in them, THEN you PC them. So they do not require sterile pouring in other words. This is LITERALLY eliminating the biggest potential for contamination.. especially when working in a SAB and not in laminar flow. I recommend them because they allow you to confidently work with agar inside of a SAB. Many can and do pour plates in SAB... but it is more challenging and risky especially when learning this hobby.

I also wish to clarify that spore --> agar is the best first step almost always. Spores are not inherently clean, in fact even when you order them from a sterile facility they are often containing small contaminants. To reduce wasted effort over time, make it a rule of thumb to always go spore --> agar .... or clone --> agar. Basically Agar is step 1 any time you are not 100% certain you have clean genetics.

Finally, test. Test. Test again. When i adjust my setup, or every so often, I will leave an agar plate OPEN in front of my flow hood for a while. Then seal it and shelf it for 2 weeks. This is WITHOUT inoculating it. This is just to make sure its sterile. If ANYTHING grows, then my technique was flawed at some point.

I also advocate testing your genetics this way over time. Until you see it grow clean on agar you cant be sure. I just had a Master Grain Liquid Culture jar go sour on me. I'm not even sure how honestly. Likely didnt flame a syringe good enough first... but I PC ALL syringes in sealed jars prior to usage in the first place.

Anyway, I didnt test the MGLC before using it... so I now have 50 contaminated agar plates as well as almost a dozen grain bags. All because I got lazy and did not do periodic controlled testing.

**I did edit a sentence to remove use of phrase "fairly devoid" as that isn't technically accurate. Also edited line regard plate refrigeration for clarities sake.

Edited by syroth (07/10/19 03:32 PM)

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Offlinejbgtaa
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Re: Workflow? [Re: syroth]
    #26100405 - 07/10/19 02:16 PM (5 years, 6 months ago)

Wait what did you say about colonizing plates in the fridge? OP, don’t do that. Allow you plates to germinate, flip them upside down, and allow them to colonize at 75 degrees Fahrenheit. THEN if you’re not going to transfer or use it to inoculate, then u can put it in the fridge. I’ll edit this post as I read the post above me to see if there’s any other incorrect information...
CVG is absolutely not devoid of nutrition, , that’s why overlay happens with too much misting... coir is completely nutritious..


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Edited by jbgtaa (07/10/19 02:19 PM)

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OfflineSoccrates
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Re: Workflow? [Re: jbgtaa]
    #26100488 - 07/10/19 02:56 PM (5 years, 6 months ago)

He said noc plates and put them in the fridge for storage, except the one you are currently using. That's right. Usually they get colonized then refrigerated, but it's the same idea


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Offlinesyroth
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Re: Workflow? [Re: jbgtaa]
    #26100566 - 07/10/19 03:31 PM (5 years, 6 months ago)

    @jbgtaa I do not appreciate your accusatory phrasing and implications. Furthermore, I've produced more information at a higher accuracy rate than your self. Most of your last comment is misleading, presumptious, and ignorant of several specific disclaimers I mentioned just because I know this is the internet and people love to quick-glance and pass quick judgement. Meanwhile, in my several written pages at this point you only found what, two single points of confusion? My usage of the phrase "fairly devoid" was a poor choice from a linguistical perspective. Although Fairly implies not all of in that usage, its still technically not correct for me to have used it in that manner.

    In other words, a poor grammatical choice is the worst mistake I've made. I am totally OK with this, I'm human, I'll make similar mistakes again.

    I never said inoculate or colonize in the fridge. Closest I see is this "Best / Longest lasting would be a thick AND extra-nutritious agar plate which, once inoculated, is promptly wrapped with parafilm and then stored at fridge temps." which I could see as being misleading to the unfamiliar.

    Cold slows growth... I kinda assumed people got that much. However in light of your confusion I have edited the post to read as follows "Best / Longest lasting would be a thick AND extra-nutritious agar plate which, once inoculated, is promptly wrapped with parafilm and then once colonized stored at fridge temps.". I was attempting to specify that you should wrap with parafilm as soon as practically possible. The refrigeration step was meant to be a logically separate judgement.

    CVG is relatively speaking devoid of nutrition, when compared to the original spawn medium. The contrast is one of many suspected factors that initiate formation of fruit. FAIR point though, devoid does literally mean completely absent which is NOT at all what I actually meant to say.

    In hindsight, the use of the word devoid was improper. Devoid means a 100% lack of something, which isn't what I meant. But lets face it, its a cool word that should be used more. And just say it out loud, it even sounds cool. I misused it in a simple attempt to use the word more in general.

    Coir itself is used as moist reptile bedding for precisely this reason as it doesnt have a ton of nutrition so it doesnt typically enable molds to begin growing on it even when moist for extended periods of time, gypsum is mostly calcium, and vermiculite is a mineral.

    I have to respectfully disagree with your assertion that CVG is completely nutritious. Your use of the phrase "completely nutritious" is just as inaccurate if not more so, than my usage of the phrase "fairly devoid".

    Many wise growers here on these forums have explained, clearly, that going from nutrient packed to not nutrient packed, lol, is part of the point. When dealing with species that prefer casing, the lack of nutrition has been clearly demonstrated to be a key factor in fruit body formation. In our case here, a non-cased mono grow, I cannot say for sure if the lack of nutrition has been scientifically linked to fruit formation as it has been in the example of casing layers, but I see a logical association there for sure.

    Regarding flipping of plates, that advice is out of place and you present it as if I was lacking such suggestion. It is OP's first monotub and they are engaging us here on a beginners level. Nuances like condensation control will take OP hours to learn the nuances of as well as several iterations of performing the work to get the hang of it. Just like we all did.

    Beyond that, I did SPECIFICALLY state several times, in fact so many that I felt silly and annoying for it, that I was leaving out details for sake of brevity, and that I was leaving out many details related to sterile procedure.

    Jbgtaa we were engaged at a high level, and you chose to incorrectly confront me on low level details that are honestly out of scope. To boot, none of the content or details you just mentioned are actually likely to have any real impact on anyone's grow.

    If your sterile procedure is GOOD, and you've tested this, and you mixed your agar RIGHT, then you can absolutely take a plate inoculate it with WAY more LC/LI than you would ever need to (lets say 2 ml for giggles, because I've done this exact thing this exact way) and then immediately wrap in parafilm and then IMMEDIATELY place in your refrigerator TOP SIDE UP.

    And guess what will happen? It will grow just fine lol. I appreciate discourse of any kind. I appreciate you pointing out my use of the word devoid. I also appreciate you letting me know I wasn't as clear as I should be (we should all be as explicit as possible when educating). Just let me be perfectly clear that I found your tone mixed with inaccuracy as unnecessary. We are clearly like minded, our process and procedure seems the same, both seem to have much experience ( I presume you do agar / culture work regularly ) and are both donating our free time voluntarily to give back to this community.

    We are not competing forces, ideally anyway. We should strive to provide synergistic and complimentary advice to those who are seeking it, and do our best to minimize semantic bickering in the form of quick replies to quickly composed commentary.

    In the same way we have to read between the lines when learning all this material, we need to continue reading between the lines as we interpret another persons communication. Typo's, rush, mis-reading, bla bla are all going to happen. You will mis-speak again, as will I. The best impact we can have over the situation is to control how we react when this happens, by choosing to do so with grace and humility.

  For example, you mis-spoke several times in your comment just now. However I have tried to understand your MESSAGE and not nit pick at what is clearly an accidental error. Specifically "Allow you plates to germinate, flip them upside down, and allow them to colonize at 75 degrees Fahrenheit. THEN if you’re not going to transfer or use it to inoculate, then u can put it in the fridge" <--- Not exactly the perfect description for that.

    However, I get that you know that, and in the context of our discussion here its too low level to warrant going back and forth to ensure perfect accuracy as its safe to assume OP will have to do much more independent learning on the subject, at which point OP will understand what you MEANT to say as opposed to how you ended up saying it.

    Another quick example in that sentence, you say 75F as if you know the species we are dealing with? We dont. Could be blue oysters, could be pan cyan for all we know. Point is, if OP is going to ever end up getting good at this then OP will certainly be able to readily recognize that although you SAID specifically 75 you clearly made a few assumptions which although well intentioned may not always be applicable to every situation. 75 is essentially room temp which is well within liveable temps for most species so I wouldn't even mention this except to highlight my point regarding communication.

Edited by syroth (07/10/19 03:53 PM)

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Offlinejbgtaa
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Re: Workflow? [Re: syroth]
    #26100663 - 07/10/19 04:19 PM (5 years, 6 months ago)

I was absolutely not trying to attack you, your grammar, or your credibility. All I was saying was not to store colonizing plates in the fridge, and they’ll need to be upside down if your incubating above 60F.

And you are correct, I don’t know which species he’s using. Op, if it’s cubes, then 75-78, and down to 70, are good colonizeng temps, for anything really.

I’m absolutely not trying to fight with you bud, it just seems as though OP is a little in over his head, and the first time I read your post there’s was conflicting info. Re reading it, it all makes a little more sense then the first read which was really just a skim.


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Edited by jbgtaa (07/10/19 04:28 PM)

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