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Re: AGAR ENVY! (Anything and All things agar!) [Re: psycho_nauticus]
    #26512948 -

7g LME / 10g agar / 500mL H2O is the standard recipe. So yeah, yours is backwards. Your agar will be pretty soft, so streak very lightly or, if you're swabbing, consider just cutting off a bit of the fiber that has spores on it and dropping it in.
Should be workable, you'll just have to do it very delicately and expect some difficulty/failure.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: psycho_nauticus]
    #26512950 -

Yeah, if your plates thin you'll see the bottom of the plate. You can still use it, just be very gentle when you do it. On harder agar it'll easily roll, soft it wants to dig in. Just be careful and I'm sure you'll be fine. you should be good psycho, both of you should. If your doing a transfer, once again just be careful, I screwed mine cause the damn transfer got stuck and I tried to use my agar to pull it off gently, now I've got a pothole but I can still transfer from it or put to grain. It's just an uggo.


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Crack's Tek's and Shit


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Re: AGAR ENVY! (Anything and All things agar!) [Re: Crackatoa]
    #26512980 -

Not as soft as 4g agar, 7g potato, 50% oat broth, 500mL LOL
That stuff is like jello, but I didn't dig in.

Anyway I'm getting on the standard agar bandwagon.
And, for extra credit I'm joining Team Telephone, 
:panic:

Is this still a practice?
Quote:
Therefore,  we  highly  recommend using  peroxidated  agar  whenever  doing  tissue  cultures,  unless  you  find  that the  species  or  strain  in  question  does  not  tolerate  it. (2006) Handbook p.114


This too
Quote:
Seal the plates with parafilm, mark them appropriately, and place them in your incubator. Store them right side up until they begin to grow out, and then reverse them as usual


Edited by Inthepit (03/02/20 07:44 PM)

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Inthepit] * 1
    #26513305 -

The only time I see a reason for antimicrobial agar is when taking a particularly difficult/dirty wild tissue clone.
Otherwise, it kind of defeats the purpose of tissue culturing - you want to be able to see everything that's present growing or else you might just keep transferring dormant/inhibited contaminants.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: footpath] * 1
    #26513324 -

footpath said:
The only time I see a reason for antimicrobial agar is when taking a particularly difficult/dirty wild tissue clone.
Otherwise, it kind of defeats the purpose of tissue culturing - you want to be able to see everything that's present growing or else you might just keep transferring dormant/inhibited contaminants.



Peroxide definitely isnt.

Flipping plates has pros and cons. I keep them right side up, they can dry out upside down

Incubation is not neccessary at room temp

I ❤ Parafilm




footpath said:
The only time I see a reason for antimicrobial agar is when taking a particularly difficult/dirty wild tissue clone.
Otherwise, it kind of defeats the purpose of tissue culturing - you want to be able to see everything that's present growing or else you might just keep transferring dormant/inhibited contaminants.


Yeah pretty much. I only uses it very rarely doe specific purposes, like you mentioned. Usually for cleaning up a VERY bacterial clone or print.

I also like to use it sometimes on the plate before I make slants (not the slant itself) to knock out any hidden bacteria before making a long term slant


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C10's Agar Guide + Tips and Tricks | c10's Flow Hood Build Guide


"Partial knowledge is more triumphant than complete knowledge; it takes things to be simpler than they are, and so makes its theory more popular and convincing."

"Convictions are more dangerous enemies of truth than lies"
― Friedrich Nietzsche

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Re: AGAR ENVY! (Anything and All things agar!) [Re: c10h12n2o]
    #26513465 -



Here's one of my neglected AA+ swab dishes that I never transferred. So cool and veiny looking

I've done 3 sets of transfers for my Puerto Rico dishes and they look terrible compared to my first swab AA+.


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TheWitchfinder's LAGM2020 Log

Edited by TheWitchfinder (03/02/20 09:13 PM)

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Re: AGAR ENVY! (Anything and All things agar!) [Re: TheWitchfinder]
    #26513470 -

Usually it does that right before they pin


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C10's Agar Guide + Tips and Tricks | c10's Flow Hood Build Guide


"Partial knowledge is more triumphant than complete knowledge; it takes things to be simpler than they are, and so makes its theory more popular and convincing."

"Convictions are more dangerous enemies of truth than lies"
― Friedrich Nietzsche

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Re: AGAR ENVY! (Anything and All things agar!) [Re: c10h12n2o]
    #26513481 -

I'll probably just take a pin and move it over. That, or drop to a grain jar for funsies. (Yes, I know that it may be full of hidden contams).

I have 10 more just like this, so we'll see. I should really just buck up and make some transfers this weekend.


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TheWitchfinder's LAGM2020 Log

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Re: AGAR ENVY! (Anything and All things agar!) [Re: TheWitchfinder]
    #26513548 -

Hey guys. How many transfers is too many?

I saved this guy from a plate used for liquid innoc.
It looks good but it’s a T6..... :shrug:


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Tek's I use

LAGM2020

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Re: AGAR ENVY! (Anything and All things agar!) [Re: staytrippy420] * 1
    #26513580 -

My mazatepec culture was on 25 and it still was ok, it started to get weird after 30, I think I had something riding on the culture so I went to spores. But I don't think there is to many transfers.


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Re: AGAR ENVY! (Anything and All things agar!) [Re: footpath]
    #26513582 -

I have sterile cutips, you think that even streaking that very softly will damage the agar? I wanted to swab and do a streak, but I guess I could just scrape off some off the print still right and drop it in?


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Re: AGAR ENVY! (Anything and All things agar!) [Re: MH5109]
    #26513583 -

MH5109 said:
My mazatepec culture was on 25 and it still was ok, it started to get weird after 30, I think I had something riding on the culture so I went to spores. But I don't think there is to many transfers.



Holy shit! haha ok cool thanks for the info man. I’ll keep it going. :rockon:


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Tek's I use

LAGM2020

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Re: AGAR ENVY! (Anything and All things agar!) [Re: psycho_nauticus]
    #26513594 -

It doesn't matter, I use a tweezers to scrape some spores up and then put it into agar. Or you could use a swab but just get it into the agar, the spores will do their thing.


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Re: AGAR ENVY! (Anything and All things agar!) [Re: staytrippy420]
    #26513599 -

If you knowingly have a contaminated syringe (Trichoderma) can I just transfer it to agar and then isolate the mycelium from there? I just prepared 10 agar plates and I am going to try unless someone recommends otherwise.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: staytrippy420] * 2
    #26513607 -

staytrippy420 said:
MH5109 said:
My mazatepec culture was on 25 and it still was ok, it started to get weird after 30, I think I had something riding on the culture so I went to spores. But I don't think there is to many transfers.



Holy shit! haha ok cool thanks for the info man. I’ll keep it going. :rockon:


I've got some on nearly 100 transfers, all good

I think the "loss of vigor" is greatly exaggerated,  especially on agar. I mean the myc is gonna expand like 1000x more than a plate does once it hits grain and bulk sub... it would be silly to say myc loses vigor once it hits grain/sub, usually it does exactly the opposite, gains vigor.

Also keep in mind that a clone is basically the same myc expanding farther and farther

So when people worry about "too many transfers" and "loss of vigor" I think it's kinda misguided.  I wouldnt worry about that until you've been dealing with the same culture for many, many years. Doesnt mean it's impossible but I think its unlikely


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

C10's Agar Guide + Tips and Tricks | c10's Flow Hood Build Guide


"Partial knowledge is more triumphant than complete knowledge; it takes things to be simpler than they are, and so makes its theory more popular and convincing."

"Convictions are more dangerous enemies of truth than lies"
― Friedrich Nietzsche

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Re: AGAR ENVY! (Anything and All things agar!) [Re: c10h12n2o]
    #26513750 -

Oh shit. I had no idea it could be stretched out that far..
Thanks for the info. I’m excited to see how far I can take these cultures.


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Tek's I use

LAGM2020

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Re: AGAR ENVY! (Anything and All things agar!) [Re: staytrippy420]
    #26514213 -

Hey guys. Was wondering what recipe people use to get the clear agar. I’ve used 8g agar and 8g BRF to 400ml of water for a long time and usually get good rhyzo growth. Would like to try something new though to get the clear agar. Let me know where I can find the ingredients to make clear agar and what y’all use to make it. Thanks in advance


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TEKs I Like

Edited by MadHatter333 (03/03/20 10:55 AM)

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Re: AGAR ENVY! (Anything and All things agar!) [Re: MadHatter333]
    #26514224 -

LME gets pretty clear...


edit:

Amazon or a brewing supply store.

Edited by footpath (03/03/20 11:02 AM)

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Re: AGAR ENVY! (Anything and All things agar!) [Re: footpath]
    #26514228 -

Cool 😎 I’ll get some of that shtuff


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TEKs I Like

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Re: AGAR ENVY! (Anything and All things agar!) [Re: MadHatter333]
    #26514238 -

MadHatter333 said:
Hey guys. Was wondering what recipe people use to get the clear agar. I’ve used 8g agar and 8g BRF to 400ml of water for a long time and usually get good rhyzo growth. Would like to try something new though to get the clear agar. Let me know where I can find the ingredients to make clear agar and what y’all use to make it. Thanks in advance


I just ordered Briess LME $9/Lb and Telephone Agar Agar from Ebay.
:kittytakedown:

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