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Offlineverum subsequentis
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an answer that turned into a thread... slants, masters and slaves, cold storage, clone hunts.... * 19
    #25911960 - 04/02/19 06:45 PM (1 year, 7 months ago)

It's all about masters and slaves. ok... I'll type it out but a little searching would yield these answers.

Let's say you have an ms tub that puts off a really nice flush with a couple of really really nice specimens mixed in it.



take those specimens



in a SAb or in the flow, with iso cleaned gloves, peel the fruit in half. You'll find it is a little odd getting started but, once peeling has begun, it runs like a banana. Be as careful as you can not to contaminate the inner stipe tissue. Nature produced it perfectly clean (unless you have such a trich problem that it's literally parasitically consuming the fruit body from the inside out).

With a flame sterilized scalpel, spoon, kitchen knife, tweezers, paper clip.... scratch a tiny piece of myc out of the inner stipe. Place this tissue on agar and wrap with glad cling wrap or parafilm. Label the petri appropriately with variety and what not. I like to write down the particular qualities that I'm hoping to replicate as well. I.E "PE, 150 gram cluster, 79 gram fruit, thick solid stipe belled cap...." obviously, you can only write so much on a petri so sometimes the petri will just say "CL-23" and i can look up all the details in my clone section of one of my many MAGIC notebooks.



If done correctly, this will be clean on the first plate. If not, make small careful transfers of the fastest, most organized, cleanest myc from each clone to a new plate. Do so until clean. At this point you go to grain with however many clones you are hoping to test out. When you do this you'll take a transfer from the plate to a new plate (more on this plate later) and put the rest of the plate to a quart (or two or three or four) of grain. Label appropriately.

Set them in a relatively temperature stabilized environment around room temp/ comfortable temp/ 60 to 75 F. Let them chill until 20-40% colonized and then shake.



Set aside again and wait for 100% colonization.



Without writing up an entire "how to do everything tek" I'll just say, make up enough coir to test all clones. I like to test clones in shoe boxes with one quart spawn and three quarts coir. It's important to test all clones in very similar environments with very similar ingredients and conditions, as to get an accurate feel for how they do compared to one another.



Ok, back to that plate we made from the clean clone plate that went to grain. When that sucker is about as big as a quarter, I wrap it in foil really well. I really like to be as clean as possible when doing this. Temperature fluctuations that are inevitably to come will try as hard as possible to suck nasty shit into this dish. When I'm feeling up to snuff and determined to be excellent I'll do this all in the FLOW or in a sab. Iso everything... gloves, arms, dishes, table, and glad wrap. Pull off a foot or two of foil and discard. Apply an extra wrap or three to each dish and wrap nicely in foil. I then place them in freshly opened gallon zip lock storage bags. I like to label the outside of the bag with exactly what cultures are in each foil wrap. Then throw those suckers in the myco fridge and let em chill until you find out what's trash and what's gold.

Then you wait while keeping good conditions on all your clone tubs and see what you get.




Harvest and weigh everything. Takes notes on everything, colonization times, wet weight, dry weight, ease of harvest... You'll then know what is worth keeping.

Finally, on to the masters and slaves. Pull out all the cultures from the fridge and allow to come back to temp while still foiled and ziplocked. Overnight is best. In the meantime you can make up your slants. I use falcom centrifuge tubes for multiple reasons. Mainly because they are cheap and easy to work with (because of the large mouth). Hamloaf turned me onto them.

Here is his write up on making slants with falcon centrifuge tubes

I follow it pretty much straight up. I think he recommends a different agar/ nute ratio for slants but I just use 2% always. Also, if using these centrifuge slants, make absolutely certain that you don't have the lids on tight and don't exceed 15 psi. I melted the ever living balls out of my first batch.

Now in a sab or flow, iso cleaning everything as normal, you'll open up your plates from the cold storage ziplock and foil, set aside the cultures that proved to be worthless little fuckers and place all winners in line for processing.

Now, i like things the way i like them and you may not, but I do this with each plate that i want to save. First take a small transfer and place it in your slant. Screw the cap on and wrap it before proceeding. In my culture work I always do the most important thing first. The slant is most important in this case so i take care of it, label it and set it aside. 

I then make two plates of the culture. One master and one slave. The slant and master plate  get grown out 2/3(ish) of the way and placed (properly) in cold storage. I let the slant grow out all the way actually. It makes it a little easier to reach in and get some later if it's grown most of the way to the top.

The point of this whole deal is this, You get your proven genetics into slants as quickly as possible to circumvent senescence. The slant can live for years in cold storage with almost NO consequences of aging and only needs taken out whenever the master plate starts showing signs of aging.

Here is where you do things however you personally find to be best. I use LC. So the slave plate makes up a 1000 cc LC and goes back in cold storage. That LC does whatever it does .




(a lot... Keep in mind, i also help manage a shiitake farm) and I only have to bust the slave out when i need a little more myc. I can use this slave until it runs out of myc and then make a new slave plate from it. Keeping track of how many slaves deep you are you'll end up learning how far that plate can expand until it starts loosing vigor. I only need to bust the master out when the slave either contams or looses vigor. Then you bust out the master plate and make a new slave. This slave is theoretically just as young and vigorous as the master plate. The master would then go back in cold storage.

You just reset everything to the age of the myc when it went into cold storage originally. If you carry on with this you'll eventually run out of myc in your master plate. So you'd take it out and make a new master with it and label it Master P2 (master plate 2.... "MP2" It's all about understandable shorthand when it comes to labeling cultures. "PE-42-MP2" tells me everything i need to know.)

Keeping track of what master plate you are on, you will finally learn that even the master got old. Or you fucked up somewhere along the way and now it's a bacterial green forest. Not to worry, the master slant waits patiently. Always treat the master slant like motha fudgin GOLD. It's the last line of defense against you fucking up and loosing everything. It's even cool to have a master slant and a slave slant if you aren't competent enough to repeatably draw from the master over the course of a couple years without pooping in it.

ummm.... So, It's all about culture age or cell expansion. That's why we die, right. a tiny little human turns into a big human and we are now old. It's far more complicated than that but ... you get the point. The master slant is like a teenage you, cryogenically frozen and able to be tapped into at any moment.

You may choose to expand with grain to grain transfers. Most are a little afraid of LC but i aint skeered. You just have to be careful with liquids or they'll bite you right in the ass. Grain to grain is the most common method of mycelial expansion. One plate makes four quarts (easily) Four quarts g2g into forty (easily) and forty quarts become 400 (easily).... and so on and so on. I've never tested the limits of g2g expansion but typically don't like to push it for a few reasons. Firstly, who the fuck needs 4000 quarts of spawn? Secondly, every G2g session is another possible fuck up. Each new round is a step further away from the original plate. None of us are perfect and the less potential vectors the better.... Alright, this is by far my longest response to any post ever. In fact, maybe I'll just make a thread of it. No proof reading or anything. My brain is tired.


PEACE.


P.S. I finally went back and "proof read". I hate typos. They make me feel dumb.


Edited by verum subsequentis (05/07/19 12:03 AM)


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InvisibleBoogieman47
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Re: an answer that turned into a post. slants, masters and slaves, cold storage, clone hunts.... [Re: verum subsequentis]
    #25911977 - 04/02/19 06:54 PM (1 year, 7 months ago)

Those are some bad ass genetics man pe are the shit nice write up... I miss seeing those tubs blow up Haha.. soon real soon :justcantwait:


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InvisibleGypsyCurse


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Re: an answer that turned into a post. slants, masters and slaves, cold storage, clone hunts.... [Re: verum subsequentis] * 1
    #25911988 - 04/02/19 06:58 PM (1 year, 7 months ago)

Wow...:takingnotes:
And thers a girl, and a shroom the size of a beer!


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Offlinepoisoned
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Re: an answer that turned into a post. slants, masters and slaves, cold storage, clone hunts.... [Re: GypsyCurse]
    #25912023 - 04/02/19 07:17 PM (1 year, 7 months ago)

Nice writeup!


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OfflineLuminous7
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Re: an answer that turned into a post. slants, masters and slaves, cold storage, clone hunts.... [Re: poisoned]
    #25912072 - 04/02/19 07:44 PM (1 year, 7 months ago)

Sweet that was helpful thanks.


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OfflineRomeoPapa
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Re: an answer that turned into a post. slants, masters and slaves, cold storage, clone hunts.... [Re: Luminous7]
    #25912130 - 04/02/19 08:24 PM (1 year, 7 months ago)

Sweet write up Verum! Much respect. :toast:


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OfflineNosmoKing
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Re: an answer that turned into a post. slants, masters and slaves, cold storage, clone hunts.... [Re: GypsyCurse]
    #25912142 - 04/02/19 08:33 PM (1 year, 7 months ago)

Beer, babes, giant shrooms and great tek, what a thread!  Thanks.


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OfflineLuminous7
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Re: an answer that turned into a post. slants, masters and slaves, cold storage, clone hunts.... [Re: NosmoKing]
    #25912228 - 04/02/19 09:27 PM (1 year, 7 months ago)

WHat is the 42 for? in  "PE-42-MP2"


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Offlineverum subsequentis
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Re: an answer that turned into a post. slants, masters and slaves, cold storage, clone hunts.... [Re: Luminous7]
    #25912233 - 04/02/19 09:32 PM (1 year, 7 months ago)

that'd be the culture number that corresponds with the Notes in my magic notebood. It would technically mean that i'm at least 42 cultures deep in PE. Sounds crazy but i'm more like 100 PE cultures deep.

to break that particular label down, it would mean, Penis Envy, culutre # 42 (see notes on culture), master plate two.


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Re: an answer that turned into a post. slants, masters and slaves, cold storage, clone hunts.... [Re: verum subsequentis]
    #25912238 - 04/02/19 09:36 PM (1 year, 7 months ago)

That's how label my clones pretty much Haha


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OfflineGrimsweeper
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Re: an answer that turned into a post. slants, masters and slaves, cold storage, clone hunts.... [Re: Luminous7]
    #25912239 - 04/02/19 09:37 PM (1 year, 7 months ago)

Nice write up VS. I started out storing my cultures in LC's but I don't like working with them. Slants are the bomb. I'm Lucky enough to have a very vigorous sporeless oyster culture in a slant Ill be able to work with for years now.


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Offlineverum subsequentis
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Re: an answer that turned into a post. slants, masters and slaves, cold storage, clone hunts.... [Re: Grimsweeper]
    #25912246 - 04/02/19 09:41 PM (1 year, 7 months ago)

Sporeless oysters? Damn... That's cool.

Thanks for the kind words everyone. I was just a few posts into answering some questions in some thread and got tired of all the half answers that make searching for info so hard. So i decided to give this particular dude a nice thorough answer. By the time i was done i thought it'd be better to make a thread out of it so more people can find the answers they need, in one place, when they search, "clones" or "slants".

I'll probably add to it over time but... we'll see.


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OfflineLuminous7
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Re: an answer that turned into a post. slants, masters and slaves, cold storage, clone hunts.... [Re: verum subsequentis]
    #25912253 - 04/02/19 09:46 PM (1 year, 7 months ago)

Quote:

verum subsequentis said:
that'd be the culture number that corresponds with the Notes in my magic notebood. It would technically mean that i'm at least 42 cultures deep in PE. Sounds crazy but i'm more like 100 PE cultures deep.

to break that particular label down, it would mean, Penis Envy, culutre # 42 (see notes on culture), master plate two.



So 42 cultures means you have taken cultures from the master slant 42 times?


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Re: an answer that turned into a post. slants, masters and slaves, cold storage, clone hunts.... [Re: Luminous7]
    #25912255 - 04/02/19 09:47 PM (1 year, 7 months ago)

No 42 different mushroom clones


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OfflineLuminous7
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Re: an answer that turned into a post. slants, masters and slaves, cold storage, clone hunts.... [Re: Boogieman47]
    #25912257 - 04/02/19 09:47 PM (1 year, 7 months ago)

Quote:

Boogieman47 said:
No 42 different mushroom clones



Ohh gotcha..


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OfflineLuminous7
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Re: an answer that turned into a post. slants, masters and slaves, cold storage, clone hunts.... [Re: Luminous7]
    #25912259 - 04/02/19 09:48 PM (1 year, 7 months ago)

All those 42 being from the original MS tub?


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Offlineverum subsequentis
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Re: an answer that turned into a post. slants, masters and slaves, cold storage, clone hunts.... [Re: Luminous7] * 1
    #25912270 - 04/02/19 09:56 PM (1 year, 7 months ago)

yep. First culture would be 1. sometimes i take multiple sectors from the same clone and label them 1a 1b 1c... You know PE-32a....

and no. not from the same tub. labeling can get a little more intense. like "PE-BO-CL-C-F3-3/21/19" that'd mean pesa boner clone C generation 3 germination date 3/21/19. that's one of my favorite cultures at the  moment.


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OfflineLuminous7
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Re: an answer that turned into a post. slants, masters and slaves, cold storage, clone hunts.... [Re: verum subsequentis]
    #25912275 - 04/02/19 10:01 PM (1 year, 7 months ago)

Quote:

verum subsequentis said:
yep. First culture would be 1. sometimes i take multiple sectors from the same clone and label them 1a 1b 1c... You know PE-32a....

and no. not from the same tub. labeling can get a little more intense. like "PE-BO-CL-C-F3-3/21/19" that'd mean pesa boner clone C generation 3 germination date 3/21/19. that's one of my favorite cultures at the  moment.



Damn son lol thats intense. And that wall of petris holy fuq dawg. This makes me excited. I love creating magical journals and keeping things really organised, my memory is shit so i log EVERYTHING. Cant wait to start preserving genetics lineages !  Thanks again.


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Re: an answer that turned into a post. slants, masters and slaves, cold storage, clone hunts.... [Re: verum subsequentis]
    #25912276 - 04/02/19 10:03 PM (1 year, 7 months ago)

I dont get that detailed as that I will just label pe1 whether its 5 transfers from that clone or not I usually always transfer atleast 3 times a plate I'd run out of room


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Re: an answer that turned into a post. slants, masters and slaves, cold storage, clone hunts.... [Re: verum subsequentis]
    #25912280 - 04/02/19 10:06 PM (1 year, 7 months ago)

Love this write up :laugh:


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