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Re: AGAR ENVY! (Anything and All things agar!) [Re: blazedup]
    #27870926 -

Yes… and no lol. Watch how much your stretching the parafilm. Starting out, I kept my wrapped plates with transfers and LC test stacked in a shelf. I would pick them up and examine them often. They were fine, but then after a week little holes developed long the film, letting stuff to get sucked in from picking up and putting down. I was getting growth along the very edges of the plate, and I’d look close and there’s a tiny hole that was exactly in line with the spot of contamination.
Maybe I got a bad batch of parafilm, but now I cut a double wide piece and fold in half before wrapping. So there’s a double layer and it can be stretched without worry. Then I put them in a ziplock bag. I’ve had zero problems since.



Edited by Seamonkey84 (07/20/22 08:58 PM)

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Seamonkey84] * 1
    #27870932 -

Cling wrap all day baby. I personally hate parafilm and will never go back.


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Two eyes to look, One eye to see.
Prying open my third eye

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Re: AGAR ENVY! (Anything and All things agar!) [Re: MysticMycologist] * 1
    #27871031 -

I've settled on horticultural grafting tape.  It's like thin cling wrap, already cut to 1" wide, but with the wax film Parafilm has. Make sure to get the Polyethylene or PE kind, which is oxygen permeable. Perfect width to wrap a plate, no paper backing, tears without tools.  No contam issues for me, I usually go 2 full wraps + a bit. Link below or search "Polyethylene grafting tape" on your fave shopping site.

https://www.amazon.com/CoscosX-Stretchable-Floristry-Self-Adhesive-Bio-degradable/dp/B07NCF4P9M


Edited by Shroomintune (07/20/22 11:03 PM)

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Tormato]
    #27871037 -

When doing josex biopsy method to clone to a lc should I lift the lid or ship?  Not sure because of piece of mushroom on the needle. Thanks


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BEWARE OF PICK POCKETS AND LOOSE WOMEN!!
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Re: AGAR ENVY! (Anything and All things agar!) [Re: wild_aussie] * 1
    #27871400 -

Good conversation, you guys. I'm also back to agar work in the last few months after a long time away. Things change. There's obviously more creativity, interest and pride in quality of work (people are growing beautiful plates using food coloring, for example--I didn't see that 10 years ago), and the knowledge level seems broader now and more accessible with more people doing this and sharing their experiences. I really enjoy learning things here.

So... exactly as you guys were saying, I've had trich contaminate a few plates at the very edges. I wrapped them in front of the flow hood, but I don't think I was mindful enough with the parafilm, and the contams might have gotten in there later. They weren't visible until after a week or so, and I had to pitch some otherwise pretty looking plates (which causes me psychological pain lol). Maybe I stretched the film too thin or too carelessly and left an opening. I fold the parafilm over lengthwise to double it up (I didn't do that 10 yrs ago), and I am now more careful to make sure the side that is stuck to the wax paper is peeled back in front of the flowhood and that that clean side is the side I stick to the plates. No more plate edge contams since being more mindful and careful like this.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: CowsPoopShrooms] * 1
    #27871448 -

CowsPoopShrooms said:
Gotta ask since I'm still learning: What makes you think they are enmeshed with something? The thinner areas?



just because it's not uniform all the way round, like this plate from a sister transfer (ignore the bacterial blotch :smile:


I think a lot of agar work for me relies on gut feelings. I think I'm gonna pop it on a little bit of grain, see what occurs. it's a side project anyways, but always learning :laugh:


So tell me Rumfor69, what do you take a sector to be? I'm gonna do a bit of reading but do you think it's goo/ bad/ contamed/ natural?

cheers all! :awedance:

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Re: AGAR ENVY! (Anything and All things agar!) [Re: keeno]
    #27871482 -

keeno said:
So tell me Rumfor69, what do you take a sector to be? I'm gonna do a bit of reading but do you think it's goo/ bad/ contamed/ natural?




So my understanding is sectoring is the
different genetic varieties that are in
favor of working together and getting all
entangled & symbiotic. This same "working
together" is the reason a "monoculture"
would actually be a way more extreme
isolation than we think.

But getting a culture to where it's not
sectoring a bunch is good so that there's
not a big gentic battle going on in the
tub. Making it a sorta axenic gentic
community rather then a bunch of different
ones. Would also lead to more uniform
fruit size, potency, immunity, all sorts
of different stuffs.

Contamination can force genetic changes
in cultures and make them work together
differently to form sectors. Sectors
dont mean contamination I would ever say
cause the mycelium with exibit other
clearer signs it's dealing with something
that are easy to detect depending if
your dish is made right and it can be
viewed well etc

I could be wrong on my understanding and
will probably be corrected here hopefully
:badadvice:


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Links For Beginners And Beyond:mspoil:

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Guerrilla] * 1
    #27871486 -

I think a lot of agar


Edited by CowsPoopShrooms (10/14/22 06:02 PM)

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Re: AGAR ENVY! (Anything and All things agar!) [Re: CowsPoopShrooms]
    #27871598 -

How are these looking? its been over 10 transfers and i feel like i still cant get a clean plate.


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Re: AGAR ENVY! (Anything and All things agar!) [Re: shroomyg123]
    #27871633 -

shroomyg123 said:
How are these looking? its been over 10 transfers and i feel like i still cant get a clean plate.





That first plate looks a but concerning to me,. Don't know about the other 2..

I don't have a ton of agar experience, but I've had plates that looked like that and were enmeshed with something that I couldn't get rid of no matter how hard I tried.. ended up getting plate pins from it that also carried the contam.. planning on trying to get a tissue sample from inside one of the pins to see if I can save the culture that way :shrug:

I'd wait for another opinion, but it definitely looks a lot like what I've been dealing with

Best of luck :mushroom2:

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Rotnpins]
    #27871658 -

The bloby ropy look it has seems
unorganized in an unhealthy fashion.

I'd do some more transfers and stay
away from transferring any thicker
blobness. Try transfering a tiny piece
of something organized and uniform.


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

Links For Beginners And Beyond:mspoil:

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Re: AGAR ENVY! (Anything and All things agar!) [Re: rumfor69]
    #27871664 -

rumfor69 said:
The bloby ropy look it has seems
unorganized in an unhealthy fashion.

I'd do some more transfers and stay
away from transferring any thicker
blobness. Try transfering a tiny piece
of something organized and uniform.



:whathesaid: also, around 6 o'clock on the first one looks sketchy.. it has has grayish tint to it. I'd avoid that area

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Re: AGAR ENVY! (Anything and All things agar!) [Re: shroomyg123]
    #27871798 -

shroomyg123 said:
How are these looking? its been over 10 transfers and i feel like i still cant get a clean plate.





Like someone said, try taking smaller transfers. Your wedges are pretty big and something is hitching a ride. If you can’t get it cleaned up after two or three more transfers then I’d just start over


--------------------
Mehhh



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Re: AGAR ENVY! (Anything and All things agar!) [Re: Wall.E] * 1
    #27872080 -

little Texas dicks



damn condensation :mad2: had it in a fridge of a while
gotta transfer these dudes

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Re: AGAR ENVY! (Anything and All things agar!) [Re: nndmt]
    #27872823 -

T3 clone plate that's in line for testing



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Links For Beginners And Beyond:mspoil:

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Re: AGAR ENVY! (Anything and All things agar!) [Re: rumfor69] * 3
    #27872900 -

rumfor69 said:
keeno said:
So tell me Rumfor69, what do you take a sector to be? I'm gonna do a bit of reading but do you think it's goo/ bad/ contamed/ natural?




So my understanding is sectoring is the
different genetic varieties that are in
favor of working together and getting all
entangled & symbiotic. This same "working
together" is the reason a "monoculture"
would actually be a way more extreme
isolation than we think.

But getting a culture to where it's not
sectoring a bunch is good so that there's
not a big gentic battle going on in the
tub. Making it a sorta axenic gentic
community rather then a bunch of different
ones. Would also lead to more uniform
fruit size, potency, immunity, all sorts
of different stuffs.

Contamination can force genetic changes
in cultures and make them work together
differently to form sectors. Sectors
dont mean contamination I would ever say
cause the mycelium with exibit other
clearer signs it's dealing with something
that are easy to detect depending if
your dish is made right and it can be
viewed well etc

I could be wrong on my understanding and
will probably be corrected here hopefully
:badadvice:




There will always be some sectoring. Some TC here (anyone know the link?) did an experiment where they tried to isolate clone mono sectors down to like the 80th generation, but new sectors still kept forming.

Sometimes a clone culture with a few sectors is better than a clone sector with only one. Can be more potent, more viable and likely be more resistant to senescence over time. 

I believe the thing to do is this: Don't worry so much about getting single pure sectors. Grow from spore prints or swabs via agar.

Then, and this is important, EAT SOME MUSHROOMS you grew.

The ones that are strong (and maybe have some other pheno you like), clone to agar.

Transfer to just T1 or T2 to get clean spawn. Don't worry about isolating single strand.

Send to grain. Keep some of the culture in the fridge. If the shrooms grown from that are strong (and maybe have some other pheno you like), keep growing from that culture.

(and / or try to select over 6 or 8 generations via spores to get spores that have the pheno and can be shared, but that's a different story and not part of cloning.) 

I think a lot of people on here spend way too much time trying to isolate single strands of mono-culture instead of growing good strong shroom cultures...without understanding why they're doing it.

Other than first initial transfers to get clean spawn, don't bother to clone / isolate anything you haven't eaten. If you do, you might be putting a lot of time and energy into isolating the perfect shroom that doesn't get you high.

Edited by nektar61 (07/22/22 01:32 PM)

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Tormato] * 1
    #27873809 -

I've never personally seen mycelium this aggressive. This Phobos plate was really dirty, there was a spot of mold at 7 o'clock, one at 9 o'clock, and a really big molded area at 12 o'clock.. now tthe mycelium has overtaken 2 of the contaminated areas and is now overtaking the 3rd



I used the plate to cool my tools, then left it open in my SAB afterwards for a while and bumped the table a couple times, then I used the plate because it was the last one I had :laugh2:

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Tormato] * 1
    #27874212 -

Spores poppin' off at 3 days. I glanced at them yesterday and saw growth but waited a day to take pix that were easier to see.
This will be F3 of the tall x7x xNippleX I'm working on. I have 9 plates all showing growth. The life force is strong in these.



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Re: AGAR ENVY! (Anything and All things agar!) [Re: Rotnpins]
    #27874405 -

Rotnpins said:
I've never personally seen mycelium this aggressive. This Phobos plate was really dirty, there was a spot of mold at 7 o'clock, one at 9 o'clock, and a really big molded area at 12 o'clock.. now tthe mycelium has overtaken 2 of the contaminated areas and is now overtaking the 3rd



I used the plate to cool my tools, then left it open in my SAB afterwards for a while and bumped the table a couple times, then I used the plate because it was the last one I had :laugh2:



Do you wipe the carbon burntness off the blade or loop or just let it be there when cutting agar, syringing and looping? I didn't know if the burntness was a potential contamination?

Thanks for your time,

:mushroom2:


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


Mushroom Cultivation Techniques...

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Grogu] * 3
    #27874421 -

The burnt carbon flecks hurt nothing
they've been flame sterilized. Wiping
the blade afterwards with anything would
contaminate the blade.


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

Links For Beginners And Beyond:mspoil:

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