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Offlinetripluv
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first agar work - what to transfer? * 1
    #19149708 - 11/17/13 01:10 PM (10 years, 3 months ago)

On 11/6 I cloned some tissue. I made the first transfer 5 days ago on 11/12.  I have 3 dishes, but most of the myc is more tomentose than rhizomorphic.  What parts do you suggest that I transfer?

Dish #1 - I'm thinking top right section of plate at 8 oclock position.


Dish #2 - all the myc on this plate looks too fluffy to me?


Dish #3 - the top section of the plate has some good looking myc t the 12oclock position, but should I avoid that section because of the satellite colony? The right and bottom sections also look like I could use some myc from those.



I plan on just the one more transfer from these plates and growing them out.


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Re: first agar work - what to transfer? [Re: tripluv]
    #19149748 - 11/17/13 01:20 PM (10 years, 3 months ago)

you can transfer that tomentose mycellium you know. If you have rhizo then transfer that but the tomentose isn't bad.

How many new dishes do you have. Might as well transfer as much as you can. Pick the strongest healthiest fastest growing parts and the rhizo form mycellium first. Anything that's furthest away from the center is faster colonizing.

If you only have 5 dishes that have 4 wells pick the 20 best looking spots on these dishes and transfer away.


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Re: first agar work - what to transfer? [Re: bodhisatta]
    #19150117 - 11/17/13 02:47 PM (10 years, 3 months ago)

for some reason, when I clone from fresh tissue from an ms, the plates always look cottony:





Like bodhi said, transfer the fastest growing from the outer edge.  mycelium goes through amazing transformations, so u never know what u will find.



--------------------
A few thoughts on cultivation
MICROBIAL HUSBANDRY!!!!

The whole is greater than the sum of its parts


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Offlinetripluv
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Re: first agar work - what to transfer? [Re: bodhisatta]
    #19150126 - 11/17/13 02:48 PM (10 years, 3 months ago)

sounds good, thanks bodhi. any idea why the rhizo is more preferable?

thats interesting about your experience with the clone myc, blinding. that myc in the bottom pic is crazy:shocked:


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InvisiblebodhisattaMDiscordReddit
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Re: first agar work - what to transfer? [Re: tripluv]
    #19150153 - 11/17/13 02:56 PM (10 years, 3 months ago)

I don't "technically" know why, but it literally means root-form. As in it is similar to roots as a network bringing nutrients around. If I'm not mistaken, in most cases, you should start to see rhizo growth eventually after some transfers anyway, but you'll transfer the tomatose stuff first.



Or sometimes you'll get both.
It's interesting because this dish was started with a transfer of a rhizo spot on the previous dish.


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Offlinetripluv
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Re: first agar work - what to transfer? [Re: bodhisatta]
    #19150193 - 11/17/13 03:05 PM (10 years, 3 months ago)

wow that is curious.  I'll make as many transfers as I can to check out what I get.  I have another sleeve of the x plates for those transfers and I was saving a couple sleeves of monoplates for later (closer to isolate) transfers.  However, I'm being stingy with the plates cause I wont be able to buy any more for a while.  The plastic plates arent reusable, are they?


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Re: first agar work - what to transfer? [Re: tripluv]
    #19150215 - 11/17/13 03:11 PM (10 years, 3 months ago)

Nope you have to toss them, They'll melt in the PC, boo. I have glass dishes that I can pressure cook, but they're quite a hassle to work with. I've been looking into using the PP containers with the twisting lids for no pour petri dishes. I have used no-pour before with the special size 1/2 pint short and wide mason jars that come in a 4pack; however that didn't go well with condensation. The ziplocks seem to work for Violet, but I've yet to try them for myself.

I guess mycellium can go back and forth between states

Quote:

RogerRabbit said:
Mycelium will change states back and forth.  A major cause is the nutritive base of the media.  Cardboard is probably the poorest media available, thus the mycelium goes rhizomorphic as it spreads.  A more nutritious substrate such as PDA is likely to cause the mycelium to revert to a more cottony appearance.
RR




Rhizo means "root like" in plant biology terms, the mycologists borrow the word to describe mycellium
The same goes for tomentose which means "cottony looking" in plants, but describes mycellium that appears the same way.

Quote:

RogerRabbit said:
Spores from the same print are the norm.  There's plenty of genetic diversity within a sporeprint.  Rhizomorphic mycelium is strain, therefore genetic related.  When isolating strains, isolate rhizomorphic sectors.  Not all rhizomorphic sectors will be good producers, but nearly all cottony sectors will be poor producers.

Yes, rhizomorphic mycellium is a good sign that the hyphae have mated.  Monokaryons won't configure into rhizomorphs.
RR




When you pick mycelium that appears rhizomorphic then you know you're isolating something that's capable of staying rhizomorphic and a much better likelihood of being able to be a good fruiting strain.

I don't know; however if the mycelium goes back into tomentose does it loose it's "vigor" ?



Edited by Trusted cuItivator (11/17/13 03:19 PM)


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Offlinetripluv
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Re: first agar work - what to transfer? [Re: bodhisatta]
    #19150246 - 11/17/13 03:18 PM (10 years, 3 months ago)

guess i'll have to be a little stingy with em for a while which is fine since I'm just getting started with it.  Might read some more of what Violet has said about the ziplocks


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Re: first agar work - what to transfer? [Re: tripluv]
    #19150259 - 11/17/13 03:20 PM (10 years, 3 months ago)

Most people believe that the rhizomorphic mycelium is faster and more aggressive. The idea is that the strong directional movement will cover more of the available grains quicker, then fill in the gaps. That being said, speed is not the only factor in what makes good genetics and I have heard of people isolating very rhizomorphic strains only to find that they are poor fruiters in the end.

As blindingleaf said, mycelium goes through amazing transformations. Sometimes media has a big role to play. If your agar is super packed with nutes, expect to see a lot of tomentose mycelium that will change its behavior when it encounters a less nutrious food source. IME even the kind of food source can determine how it looks. I have had very tomentose mycelium on MEA, turn into crazy ropes when transfered to PDA. Of course it can change again when it goes to grains.

IMO the only time rhizo mycelium counts is on grains. I don't really care how it looks on MEA or PDA or dog food agar for that matter. That's why I like to have a few plates of grain soak water agar kicking around, I feel that that gives a better indication of how it will perform with that food source.

This is all just a bunch of anecdotal conjecture but, I say healthy growth is healthy growth no matter if it is rhyzomorphic or not. Transfer it, inoculate with it, fruit it, then decide if you like it :awesomenod:

Edit; Bohisatta beat me to the punch with some good RR quotes, nice job :super: BTW I do a lot of no pour PP5 agar and it works great for me. Twist lids are not needed either, the little containers are easy as hell to work with, can fit lots in the fridge and if they get a little loose after a few runs in the PC, its nothing a little micropore tape on the edge can't fix.



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InvisiblebodhisattaMDiscordReddit
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Re: first agar work - what to transfer? [Re: Pastywhyte]
    #19150279 - 11/17/13 03:24 PM (10 years, 3 months ago)

Violet's petri dish substitute

I would advise to experiment with the fertilizer when you're have free time accept the possible failure, or hopefully success.


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Offlinetripluv
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Re: first agar work - what to transfer? [Re: bodhisatta]
    #19150291 - 11/17/13 03:27 PM (10 years, 3 months ago)

i wont be using the fertilizer at all. MEA seems to be all purpose and all I care to use for a while mostly because I bought a bunch when I had a couple bucks.  Its interesting that I cant expect the cottony myc to produce well, but I guess its tough to tell if my MEA is really nutritious.

went right by your post, Pasty.  Really exciting about the plastic containers that can handle the PC :rockon:  How do you do the grain soak water agar??


Edited by tripluv (11/17/13 03:38 PM)


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InvisiblebodhisattaMDiscordReddit
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Re: first agar work - what to transfer? [Re: tripluv]
    #19150323 - 11/17/13 03:35 PM (10 years, 3 months ago)

Yea I just use ELME and it's been working well.

I have some of those small 1/4 pint containers pasty posted but I use them as PF test cake jars. I had a problem with a lot of them imploding in the pressure cooker unless I made a polyfil gas exchange on them. I figure if I use agar in them with the polyfil it's going to boil and saturate the polyfil and then contams will grow through it. Maybe I'll try it though.


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Re: first agar work - what to transfer? [Re: tripluv]
    #19150325 - 11/17/13 03:36 PM (10 years, 3 months ago)

I've pretty much done every variation of that for about a year now, minus the ferts. I just don't see the point in them. One of these days I'm gonna give her tek a complete shot with ferts, RGS, LED's, the works. Right now tho I just don't have the time to monkey around with everything. I barely have time for the set and forget approach of monotubs.

I have been doing no pour in glad mini rounds for about 2 years, ever since I saw spongiforms cake tek with them. I was like "I can put these in the PC?" Haven't looked back. They are a nice size, I can fit shitloads of em in the fridge, no screw top is easy in the SAB, reusable, cheap as hell, buy em in the grocery store, etc. Don't get me wrong, I do like the pour plates as well, just got in some x plates like the OP used and looking forward to getting those going, but for a cheap reliable, easy to find alternative, mini rounds work pretty good for me.

Quote:

tripluv said:
i wont be using the fertilizer at all. MEA seems to be all purpose and all I care to use for a while




You should change your media up a bit if you want to keep your cultures around. Forget the ferts but PDA, PDYA, dogfood, oatmeal, mixing it up with honey or karo, etc, is not a bad idea to hold off senescence. I usually do PDA, grain water, honey, dog food, for most of my transfers and save the MEA for my slants.


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Re: first agar work - what to transfer? [Re: Pastywhyte]
    #19150341 - 11/17/13 03:41 PM (10 years, 3 months ago)

Do you have problems with implosions using those 1/4pint rounds? Any lid modification?

Here's my first agar work thread if you want to follow along


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Re: first agar work - what to transfer? [Re: bodhisatta]
    #19150342 - 11/17/13 03:41 PM (10 years, 3 months ago)

Quote:

bodhisatta said:
Yea I just use ELME and it's been working well.

I have some of those small 1/4 pint containers pasty posted but I use them as PF test cake jars. I had a problem with a lot of them imploding in the pressure cooker unless I made a polyfil gas exchange on them. I figure if I use agar in them with the polyfil it's going to boil and saturate the polyfil and then contams will grow through it. Maybe I'll try it though.




The trick to the minis is to have a 1/4" hole for GE and a strip or two of micropore over it. I wouldn't trust polyfil. Will keep the contams out and give just enough to keep the thing from being crushed. The other thing is to bring the pressure up slowly. Bring it up too fast and it will not only crush the container, but also boil the agar up the sides. See how clean the sides of mine are? If they don't look like that, you brought the pressure up too quick.


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Re: first agar work - what to transfer? [Re: Pastywhyte]
    #19150350 - 11/17/13 03:43 PM (10 years, 3 months ago)

well I'll have to look more into the PDA and others, then...

I edited the post above...How do you do the grain water agar?


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Re: first agar work - what to transfer? [Re: tripluv]
    #19150354 - 11/17/13 03:43 PM (10 years, 3 months ago)

Quote:

tripluv said:
How do you do the grain soak water agar??




Simple, just save some of the water from the grain soak, run it through a coffee filter to get the gypsum out (assuming you use gypsum, I do) then add your agar, PC and serve :wink: Nothing to it.


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Re: first agar work - what to transfer? [Re: Pastywhyte]
    #19150364 - 11/17/13 03:45 PM (10 years, 3 months ago)

Looking good tripluv :thumbup:


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Re: first agar work - what to transfer? [Re: tripluv]
    #19150387 - 11/17/13 03:50 PM (10 years, 3 months ago)

There are a few other tricks to getting good results from the pp5 no pour but I'm not going into them now, Anyone interested send me a :pm: otherwise ya'll just have to wait for the tek to be finished :wink:


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Re: first agar work - what to transfer? [Re: Pastywhyte]
    #19150394 - 11/17/13 03:51 PM (10 years, 3 months ago)

Quote:

Pastywhyte said:
Quote:

bodhisatta said:
Yea I just use ELME and it's been working well.

I have some of those small 1/4 pint containers pasty posted but I use them as PF test cake jars. I had a problem with a lot of them imploding in the pressure cooker unless I made a polyfil gas exchange on them. I figure if I use agar in them with the polyfil it's going to boil and saturate the polyfil and then contams will grow through it. Maybe I'll try it though.




The trick to the minis is to have a 1/4" hole for GE and a strip or two of micropore over it. I wouldn't trust polyfil. Will keep the contams out and give just enough to keep the thing from being crushed. The other thing is to bring the pressure up slowly. Bring it up too fast and it will not only crush the container, but also boil the agar up the sides. See how clean the sides of mine are? If they don't look like that, you brought the pressure up too quick.




I don't trust micropore tape haha.
I use polyfill on all my grain jars and my 1/4pint PF cakes with no issues so far. I'll keep the pressure thing in mind though, and maybe try micropore since it's working for you.

Frank's agar journal w/ grain dunk info


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