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OfflinePrimalSoup
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Re: 1st experience with agar. Is this supposed to be easy? [Re: spacechildo]
    #21534379 - 04/11/15 06:27 PM (9 years, 1 month ago)

Quote:

spacechildo said:
you want to transfer healthy myc away from all the other shit that is growing!




like QFT a thousand times over...  I just can't stand to look at those pics again...


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OfflineOrganic_Magic
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Re: 1st experience with agar. Is this supposed to be easy? [Re: PrimalSoup]
    #21534438 - 04/11/15 06:44 PM (9 years, 1 month ago)

To me at least, it kind of all looks like mycelium. Two drops will spread out across a plate as soon as you mess with it so its not surprising that there's so many inoculation points.


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:pipesmoke:Its all fun and games until mushrooms make you gay:pipesmoke:




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Offlinespacechildo
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Re: 1st experience with agar. Is this supposed to be easy? [Re: DinkinFlicka84]
    #21534460 - 04/11/15 06:48 PM (9 years, 1 month ago)

its very hard to tell what is clean and what is not on those plate cause you got so much going on.
just take a transfer from a small white piece and see what grows out,
keep transferring and you'll hopefully end up with some clean myc on a fresh plate!

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InvisibleTheEaglesGift
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Re: 1st experience with agar. Is this supposed to be easy? [Re: spacechildo]
    #21534823 - 04/11/15 08:17 PM (9 years, 1 month ago)

You need to get some parafilm to wrap those dish edges, or at the very least some saran wrap. That tape is NOT doing the job properly, as you can tell by the huge gaps where it isn't even making contact with the plate.

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OfflineDinkinFlicka84
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Re: 1st experience with agar. Is this supposed to be easy? [Re: TheEaglesGift]
    #21538806 - 04/12/15 06:38 PM (9 years, 1 month ago)

I can use definitely use saran or parafilm on my next tranfers. I just happened to have micropore tape at the time. They still have no contaminants in the dishes, so it's worked so far.

Thanks for the info on the mycelium. I will cut from what looks best to me.


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OfflineDinkinFlicka84
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Re: 1st experience with agar. Is this supposed to be easy? [Re: DinkinFlicka84]
    #21544595 - 04/13/15 11:28 PM (9 years, 1 month ago)

Ok, so I did my first transfer. It was a little sloppy with cutting out the best looking mycelium, but I think I chose the whitest and most rhizopmorphic  looking spots. Just to be sure I did it right, I cut out a TINY section of a sector (~1cm long) that looked more solid white and had "legs." Is this correct? There were probably 20 spots to choose from.

However, I didn't notice any mold. Does the mold look more or less like the mycelium? White and fuzzy? Or is it discolored?

In RR's videos, he cuts 3 slits in the  receiving petri before adding  the colonized agar to it. What's the purpose of the 3 slits? I did the slits just because he did them and I tried to place the agar piece on the area of slits.

Please help me understand the point of this and if mold is hard to spot. Thanks!


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OfflineOrganic_Magic
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Re: 1st experience with agar. Is this supposed to be easy? [Re: DinkinFlicka84]
    #21544607 - 04/13/15 11:33 PM (9 years, 1 month ago)

Mold becomes easier to spot once you observe it a few times. Making the three slits is to cool the freshly flame sterilized scalpel before you make your transfer.

Its all pretty much trial and error until you acquire an eye for it. :super:


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:pipesmoke:Its all fun and games until mushrooms make you gay:pipesmoke:




            click and learn.

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OfflineDinkinFlicka84
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Re: 1st experience with agar. Is this supposed to be easy? [Re: Organic_Magic]
    #21544631 - 04/13/15 11:43 PM (9 years, 1 month ago)

That makes sense. I definitely feel like a lot of what I do with dishes is learning.
I don't know how many different petris you guys normally inoculate and then transfer, but I'd like to know.

I had 4 plates with 3 segments in each, so I had 12 inoculated segments. After that, I had 4 more plates (12 segments) to transfer the best 12 cultures in IMO.

Is that enough transfers? I just wanted to give my self enough tries to get better at technique and to see which of my mycelium would turn out healthy.

How many inoculated dishes do you normally do in search of the perfect culture?


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InvisibleNumeroEno
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Re: 1st experience with agar. Is this supposed to be easy? [Re: DinkinFlicka84]
    #21544655 - 04/13/15 11:52 PM (9 years, 1 month ago)

God I've lost track, but I started pouring petris around the beginning of the year and I've gone through more than I can count. I have over 30 plates in my fridge and like 10 more incubating, and that doesn't even compare to how many used plates I've thrown out or lost to contams. There's no right number of transfers as long as you get the culture clean. After that it's a matter of strategy.


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:gd_icon:  Let it grow! Let it grow! Greatly yield!  :gd_icon:
What shall we say, shall we call it by a name
As well to count the angels dancing on a pin
Water bright as the sky from which it came
And the name is on the earth that takes it in

DOG FOOD AGAR

MY ELECTRIC INOCULATION LOOP

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OfflineDinkinFlicka84
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Re: 1st experience with agar. Is this supposed to be easy? [Re: NumeroEno]
    #21544663 - 04/13/15 11:58 PM (9 years, 1 month ago)

I guess I mean how many different cultures at first do you cut and see if they grow well in another dish? Do you start with 10 different cultures or more?


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InvisibleNumeroEno
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Re: 1st experience with agar. Is this supposed to be easy? [Re: DinkinFlicka84]
    #21544673 - 04/14/15 12:07 AM (9 years, 1 month ago)

Quote:

DinkinFlicka84 said:
I guess I mean how many different cultures at first do you cut and see if they grow well in another dish? Do you start with 10 different cultures or more?




I don't quite understand. Say you're going from a multispore inoculation on agar, you're starting with one culture, but you could take 3 transfers from that, and then make 3 more transfers from each one exponentially.

There's no right or wrong amount of transfers to take, you just transfer until you can pick out a nice sector to work with, but the criteria are pretty subjective.

Most people go for aggressive rhizomorphic growth, and since primordia form more readily on rhizomorphic growth that's a pretty good way to go. After that it's up to you whether you want to isolate further, or use younger mycelium.

I believe in keeping the mycelium younger. Also, growing out young, fresh mycelium and taking a clone from a nice cluster gets you a good fruiting set of genetics pretty quickly.


--------------------

:gd_icon:  Let it grow! Let it grow! Greatly yield!  :gd_icon:
What shall we say, shall we call it by a name
As well to count the angels dancing on a pin
Water bright as the sky from which it came
And the name is on the earth that takes it in

DOG FOOD AGAR

MY ELECTRIC INOCULATION LOOP

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Invisibleuncle_rico
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Re: 1st experience with agar. Is this supposed to be easy? [Re: DinkinFlicka84]
    #21544765 - 04/14/15 12:36 AM (9 years, 1 month ago)

Quote:

DinkinFlicka84 said:






Since you have been doing GLC for a while, take a cleanly colonized grain and drop that on agar. (assuming the culture is not 'old' calendar age or cell division age)  The high nutrient grain mycelia growing out into the lower nutrient agar from one point will greatly simplify matters.  easy way to get going with agar.


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OfflinePrimalSoup
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Re: 1st experience with agar. Is this supposed to be easy? [Re: DinkinFlicka84]
    #21546227 - 04/14/15 12:22 PM (9 years, 1 month ago)

Quote:

DinkinFlicka84 said:
I guess I mean how many different cultures at first do you cut and see if they grow well in another dish? Do you start with 10 different cultures or more?





As many as it takes, whatever that turns out to be.  You're trying to isolate a strain, right?  Or are you just trying to get a clean culture?  Same answer though. :thumbup:


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OfflineDinkinFlicka84
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Re: 1st experience with agar. Is this supposed to be easy? [Re: PrimalSoup]
    #21549400 - 04/14/15 11:26 PM (9 years, 1 month ago)

It's day 2 after my 1st transfer. When do I transfer again? The culture has started to grow from that spot somewhat, but how big does it need to be before I do another transfer?

After my first inoculation, I was told to transfer right away, as soon as it was big enough to see the rhizomorphic mycelium.


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OfflineOrganic_Magic
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Re: 1st experience with agar. Is this supposed to be easy? [Re: DinkinFlicka84]
    #21549407 - 04/14/15 11:30 PM (9 years, 1 month ago)

i transfer when its the size of a penny. :super:


--------------------
:pipesmoke:Its all fun and games until mushrooms make you gay:pipesmoke:




            click and learn.

Bod's simplified cultivation methods
Got side pins?

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OfflineDinkinFlicka84
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Re: 1st experience with agar. Is this supposed to be easy? [Re: Organic_Magic]
    #21549423 - 04/14/15 11:35 PM (9 years, 1 month ago)

Awesome. Will do! I'll post pics before I transfer.


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Offlinefunghi37
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Re: 1st experience with agar. Is this supposed to be easy? [Re: DinkinFlicka84]
    #21550624 - 04/15/15 11:07 AM (9 years, 1 month ago)

I am new and I have found it way easy and less work then having to wait for colonizing jars and waiting to see if they are contaminated! I started transferring from contaminated plates shortly after my first attempt in growing! The first time I worked with Agar I had one plate out of 6 contaminate ! And I  transferred all at penny size growth!  I did the contaminated plate last to keep the rest of my plates from having any possible contact with any contamination ! I was successful on my first transfer! I will make transfers until I have a good specimen!
And yes It is easy as pie! I just try to take extra steps to be sterile every time I work!
Keep up the good work!
And I was told by a lot of successful TC's to take notes! Study and read!!!
Sorry to just jump in on this discussion!!!

Contaminated plate
Transfer # 1
1st swab from print!
Transfer from that!

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OfflineDinkinFlicka84
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Re: 1st experience with agar. Is this supposed to be easy? [Re: NumeroEno]
    #21555122 - 04/16/15 12:31 PM (9 years, 1 month ago)

I understand young mycelium, but how do I go about getting young myc from my isolated culture or monoculture? Wouldn't the isolated culture, being transferred a few times make it older myc?

2 more things...

Do I have to wait for the culture to be 100% monoculture or can I inoculate my WBS jars with a small slice of healthy rhisomorphic mycelium?

Also, if and when I have a monoculture, do I need to duplicate that culture in another petri to have more mycelium slices for inoculating jars?

Lastly what's the best and easiest way to store the master culture? RR has test tubes and centrifical force machine that spins the test tubes. I don't have, nor do I want to spend that kind of money.


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OfflineDinkinFlicka84
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Re: 1st experience with agar. Is this supposed to be easy? [Re: funghi37]
    #21555144 - 04/16/15 12:37 PM (9 years, 1 month ago)

Funghi37, what is the swab from print all about?


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Offlinespacechildo
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Re: 1st experience with agar. Is this supposed to be easy? [Re: DinkinFlicka84]
    #21555222 - 04/16/15 12:51 PM (9 years, 1 month ago)

transferring a culture through a couple plates isn't much, putting it through fruiting and take a late flush shroom to agar is getting old-ish.

you dont need an isolate to fruit it, ms grows are fine as long as the myc is healthy (all cube myc, no mold, bacteria etc).
rhizo/tomentose doesnt matter.

if/when you get an isolate you want it on another petri so you can fruit your iso and keep it too.
store it on slanted test tubes, no need for a centrifugal force machine.
you can also store it on agar plates :shrug:

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