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InvisiblebodhisattaMDiscordReddit
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Re: Pasty Agar Tek [Re: blindingleaf]
    #20869223 - 11/21/14 07:36 AM (9 years, 2 months ago)

no transfer before it reaches the edge of the dish one dish could inoculate 20 quarts if you take small wedges.



I would use a wedge bigger than that though


Edited by Trusted cuItivator (11/21/14 07:38 AM)


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OfflinetaGyo
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Re: Pasty Agar Tek [Re: Inocuole]
    #20869225 - 11/21/14 07:37 AM (9 years, 2 months ago)

Quote:

Inocuole said:
but before you inoculate, let a drop or two fall out the syringe to cool it




I didn't do this with my first two syringes and just used my SAB for the first time two nights ago and did it and instantly understood why. The first few pushes were literally straight air. I felt like a mycologist :]

Quote:

Inocuole said:
Also no you can inoculate with a half-colonized piece of agar and it should take over as long as your procedure is good.



Got it :thumbup:

Thanks Inocuole :]

Sounds good Bod :hippie:


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Re: Pasty Agar Tek [Re: taGyo]
    #20869231 - 11/21/14 07:39 AM (9 years, 2 months ago)

Quote:

taGyo said:
I felt like a mycologist :]





You're gettin there.  The other day I went to rate you since you were kicking ass around here and then I realized I was the second person to rate you right after you showed up. :lol:


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Re: Pasty Agar Tek [Re: Inocuole]
    #20869245 - 11/21/14 07:49 AM (9 years, 2 months ago)

Quote:

Inocuole said:

You're gettin there.  The other day I went to rate you since you were kicking ass around here and then I realized I was the second person to rate you right after you showed up. :lol:



REALLY?

WOW! I just looked! Lmao, SMALL world!

It's honestly thanks to people like you, Bod, Cron, ToxicDuck, and PW that have pretty much babysat me to this point. I'm slowly letting go of the hand and taking that tentative first step forward :lol:

Thanks again guys :]


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Re: Pasty Agar Tek [Re: Inocuole]
    #20869744 - 11/21/14 10:39 AM (9 years, 2 months ago)

Quote:

Inocuole said:
Not really, the contam can only travel in 2 dimensions so if it's far enough away from the healthy growth then you can just use a tool to grab the tinest piece of clean mycelium that you can and set it down on a new plate.  It's a good idea to be on the ball about it but there isn't necessarily a rush, depending on how bad the plate is.



Thanks man I'm New to agar, you, pasty,and muddafucka are contributing good info.


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Invisiblestareatclouds
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Re: Pasty Agar Tek [Re: spirit_shadow]
    #20873435 - 11/22/14 03:22 AM (9 years, 2 months ago)

Sorry for the poor quality, I snapped it through my SAB. What do y'all think of these two? They're both transfers from another plate. Not sure why the one is so messed up. I'm assuming I got a few little chunks stuck on there.

Would y'all think these are clear to make LI with or should I transfer them some more? They both look pretty clean to me (although the chunk, weird one looks iffy to me), but I can still use an extra set of eyes on my agar.



Also, I'm having trouble picking up the entire wedge when I slice the circle out. It's just lifting and tearing the area I have my blade in. Is this because I'm making the plates too thin with myc?


Edited by stareatclouds (11/22/14 05:01 AM)


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OfflinePsyCLown89
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Re: Pasty Agar Tek [Re: stareatclouds]
    #20873815 - 11/22/14 08:22 AM (9 years, 2 months ago)

Quick question, if your first agar plate does not have any contams, then there is no need to transfer it to a second plate. Correct?


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InvisibleInocuole
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Re: Pasty Agar Tek [Re: PsyCLown89]
    #20873826 - 11/22/14 08:27 AM (9 years, 2 months ago)

Well, why not?  It depends on your goal with that culture.  If you just want to germinate spores to agar, get a clean culture, and put it to grain, you should still transfer once to make sure what you have is in fact clean.  If the second plate turns out clean, then by all means, do what you're going to do with it.  It also depends on what you used to inoculate the first plate.  If it's a clone or colonized grain and it's clean then I suppose you wouldn't need to transfer it, but always transfer away from spores at least once or twice.

For isolation and other stuff, obviously you might want to continue transferring until you have the results you want.


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Re: Pasty Agar Tek [Re: PsyCLown89]
    #20873830 - 11/22/14 08:28 AM (9 years, 2 months ago)

stare, i wouldn't use that first picture, but the second two look ok.  the first pic just has an odd growth pattern.  try using the back/blunt part of ur blade to cut the circle out, then kinda spatula it out, if that makes sense.

psyclown, yea u can use it if ur sure it has no contaminates, but like innocule said, i'd make at least one transfer.  germinating mold and germinating spores look really similar.


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Re: Pasty Agar Tek [Re: blindingleaf]
    #20873903 - 11/22/14 09:00 AM (9 years, 2 months ago)

Quote:

blindingleaf said:
stare, i wouldn't use that first picture, but the second two look ok.  the first pic just has an odd growth pattern.  try using the back/blunt part of ur blade to cut the circle out, then kinda spatula it out, if that makes sense.



:whathesaid: I would still do another transfer on the second two.


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OfflinetaGyo
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Re: Pasty Agar Tek [Re: Inocuole]
    #20873985 - 11/22/14 09:41 AM (9 years, 2 months ago)

Quote:

Inocuole said:
It depends on your goal with that culture.



Hey Inocuole,

I had heard that lower levels of nutrition created rhizo growth. So if I altered Pasty's formula a little in a few jars to have less Karo/Flakes would I see more rhizo growth? Once transferred to a nutritious grain substrate it would switch over to tomentose though right? I'm just having a hard time finding a guaranteed way to get rhizo growth on jars since mine always colonize fast but are white and fluffly.

Pasty made a post saying just look for healthy growth and I completely agree but if I'm working with Agar I want to see rhizo at least once :]


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InvisibleInocuole
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Re: Pasty Agar Tek [Re: taGyo]
    #20874472 - 11/22/14 12:35 PM (9 years, 2 months ago)

Well it's possible that altering nutritional content could affect some of the behavior of the mycelium including but not limited to making it more rhizomorphic but I wouldn't do that while trying to isolate sectors or for agar intended as an inoculate.  That extra nutrition in there helps it jump off the agar onto the grain more quickly and that's somewhat important for my current uses, since the name of the game is still a race against the clock.


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Re: Pasty Agar Tek [Re: Inocuole]
    #20874484 - 11/22/14 12:38 PM (9 years, 2 months ago)

Exactly why I stress not to worry about rhizo, I could make even weak ass cultures appear rhizo if I wanted to. Look for healthy growth.


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Re: Pasty Agar Tek [Re: Pastywhyte]
    #20874497 - 11/22/14 12:41 PM (9 years, 2 months ago)

Quote:

Pastywhyte said:
Exactly why I stress not to worry about rhizo, I could make even weak ass cultures appear rhizo if I wanted to. Look for healthy growth.




:whathesaid:

:bongload:


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Re: Pasty Agar Tek [Re: Inocuole]
    #20874553 - 11/22/14 12:50 PM (9 years, 2 months ago)

I saved that post.

Thanks guys :hippie:


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OfflinePsyCLown89
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Re: Pasty Agar Tek [Re: taGyo]
    #20878139 - 11/23/14 07:35 AM (9 years, 2 months ago)

So I managed to get all the ingredients for the agar.

Managed to cook up some agar and mash on a gas bottle while waiting for the power to come back on. I think I might have added a bit too much agar as I forgot it would need to set.

I dumped the agar into a few containers and they were already setting so it's not smooth. I'm hoping it will break down into a liquid in the pressure cooker and then that'll be fine.

Quite eager to give this a try and see what it's like. What happens if there is too much agar? Will the myc not grow as well or?


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Re: Pasty Agar Tek [Re: PsyCLown89]
    #20878145 - 11/23/14 07:37 AM (9 years, 2 months ago)

Your plates will just be stiff. Spores generally like a little soupier agar, but my first batch was real heavy on the agar and still had some spores germ on it (but not all). If you are transferring live cultures, it shouldn't be an issue, really.


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OfflinePsyCLown89
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Re: Pasty Agar Tek [Re: Munchauzen]
    #20878499 - 11/23/14 10:00 AM (9 years, 2 months ago)

I see, I was thinking for some reason that I should get a texture / stiffness similar to Jelly (Jello) whatever you wanna call it - not jam.

I will be using spores from a syringe and directly from a spore print? Think I should maybe try and make more and do it properly this time?


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Re: Pasty Agar Tek [Re: PsyCLown89]
    #20878510 - 11/23/14 10:06 AM (9 years, 2 months ago)

It will liquefy in the PC and set again once cooled. You do want it to be the consistency of jello. I like a bit softer mix for germinating spores but it should still be a solid. If you are using a syringe the spores will already be hydrated so a firmer mix will still be fine. Just remember not to use too many spores or it will be hard to track your growth.


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Re: Pasty Agar Tek [Re: MudaFuka]
    #20878757 - 11/23/14 11:38 AM (9 years, 2 months ago)

Quote:

MudaFuka said:
It will liquefy in the PC and set again once cooled. You do want it to be the consistency of jello. I like a bit softer mix for germinating spores but it should still be a solid. If you are using a syringe the spores will already be hydrated so a firmer mix will still be fine. Just remember not to use too many spores or it will be hard to track your growth.




Perfect then! I made another batch of agar as well, used blue food colouring this time (as opposed to green for the first) and made this one a bit better. Poured it when it was liquid too and looked much better and neater :laugh:

Got 4 in the PC, 2 blue and 2 green. Then one more of each in the fridge due to a small PC :frown:

Will drop a single drop from the syringe in the middle of the agar in a green and a blue mixture and see how it differs and play around a bit - hopefully start with some grain work sometime soon - need some gypsum first though. Struggling to find :frown:

Thanks! :laugh:


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