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OfflineTrippyLivin
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Saving agar that's drying out
    #24257637 - 04/19/17 03:47 PM (7 years, 8 months ago)

So these were the first dishes I poured. Ends up I poured it too thin and it's slowly pulling in from the edges of the dish even after wrapping with parafilm and Saran Wrap. They are still quite small... so these three are on the same Petri. About a month old. Should I just take the whole one for each one and do LC or should I actually try to pick the best section? It would be just so small tho...!each ones probabaly 1cm by 1cm




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Invisiblemynakedrat
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Re: Saving agar that's drying out [Re: TrippyLivin] * 1
    #24257641 - 04/19/17 03:48 PM (7 years, 8 months ago)

Transfer some of those into more plates man

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Re: Saving agar that's drying out [Re: mynakedrat]
    #24257768 - 04/19/17 04:46 PM (7 years, 8 months ago)

:whathesaid:

Just pour some new plates and transfer. I usually make at least 3 before I put to grain


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OfflineTrippyLivin
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Re: Saving agar that's drying out [Re: mushpunx]
    #24261938 - 04/21/17 02:08 PM (7 years, 8 months ago)

What do you guys think about doing a invitro grow using PF tek in quart size containers. I've got little to no experience with this but figured it would be a good tek to use. I could take a pin that grows the highest and biggest then clone it on agar then progressing towards Isolation of a monoculture from there.

Also if you were to say use the three spots on my agar that are 1cm by 1cm each. Would you use the whole thing or just cut them in half so I can put it in 6 different containers


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Re: Saving agar that's drying out [Re: TrippyLivin]
    #24262111 - 04/21/17 03:42 PM (7 years, 8 months ago)




This is a very old picture, but that's the size wedge I like to innoxulate my jars with.
Like we keep saying, take a transfer from those plates, to new plates, and then *Grow them out* to 2/3rds of the plate.


Don't put what you have there to grain.

I wouldn't waste my time with the invitro PF tek thing. You are culturing on agar, working with grains. Do a bulk grow .


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Edited by mushpunx (04/21/17 03:45 PM)

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OfflineTrippyLivin
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Re: Saving agar that's drying out [Re: mushpunx]
    #24262284 - 04/21/17 05:26 PM (7 years, 8 months ago)

Oh I already have my Bulk grows going. I'm transferring to Bulk substrate tonight when I get home. Driving home from Canada at the moment down the east coast. The idea for vitro is to eventually create a monoculture.

Anyone have any experience with sterilization strips? I just bought 650 of them http://m.ebay.com/itm/161943638850?_mwBanner=1


--------------------
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-TrippyLivin

Check out my indoor B+ and Golden Teacher Indoor Bulk Grow!
Adding AA+ Albino Cubensis,  Corumba Brazil Cubensis, Mazatapec Cubensis, and Texas PE6 Cubensis

Starting Greenhouse Grow Bulk

Edited by TrippyLivin (04/21/17 05:30 PM)

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Re: Saving agar that's drying out [Re: TrippyLivin]
    #24262302 - 04/21/17 05:35 PM (7 years, 8 months ago)

Quote:

TrippyLivin said:
Oh I already have my Bulk grows going. I'm transferring to Bulk substrate tonight when I get home. Driving home from Canada at the moment down the east coast. The idea for vitro is to eventually create a monoculture.

Anyone have any experience with sterilization strips? I just bought 650 of them http://m.ebay.com/itm/161943638850?_mwBanner=1





Why? So you put a strip in a PC and after the cycle it changes color. Doesn't really tell you anything besides that the strip turned a color. Perhaps I'm not understanding how you plan to use these.

monocultures are a waste of time for cubes. leave that shit to professional mycologists. we get plenty of bang for our buck from clones.

Edited by Munchauzen (04/21/17 06:48 PM)

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Re: Saving agar that's drying out [Re: Munchauzen]
    #24262327 - 04/21/17 05:55 PM (7 years, 8 months ago)

Getting a monoculture is easy. Finding one that performs well is what takes a lot of work

I agree, just stick to cloning right now. Finding a really awesome clone is work enough too


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Re: Saving agar that's drying out [Re: mushpunx]
    #24262374 - 04/21/17 06:28 PM (7 years, 8 months ago)

Quote:

mushpunx said:



This is a very old picture, but that's the size wedge I like to innoxulate my jars with.
Like we keep saying, take a transfer from those plates, to new plates, and then *Grow them out* to 2/3rds of the plate.


Don't put what you have there to grain.

I wouldn't waste my time with the invitro PF tek thing. You are culturing on agar, working with grains. Do a bulk grow .




I threw in about half inch of wedge into every grain jar. :frown:

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Invisiblec10h12n2o
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Re: Saving agar that's drying out [Re: mushpunx]
    #24262392 - 04/21/17 06:40 PM (7 years, 8 months ago)

i dont know about easy

99% of the people who say they have an "isolate" almost certainly do not

serial dilutions make it much faster/easier, but starting from spores, you can easily be doing hundreds of transfers before you get an isolate

most people think they have an isolate after they transfer a sector, or have organized growth, when what they really have is a group of strains. if they were to do another 10 or so transfers they might see more sectors than they would believe

i cant tell you how many times i incorrectly thought i had an isolate, only to still be working towards it 6mo later.

hell, i have one project going where i cloned a 295g fruit nearly a year ago, and just for the hell of if have been trying to isolate from it, still working on it today.... it is astounding how many strains and strain groups can be present in a fruit, and im fascinated by the dynamics of when they distinguish themselves vs forming an organized culture

@ op, id avoid all that kind of stuff for the time being... stick to proven, reliable methods until you get a few grows under your belt

as they said, transfer those to better plates... dont get attached to plates, you will go through LOTS, thats the point

also, what you were saying about getting a monoculture would not result in a monoculture, or even close

what munch said was right on target: stick to clones, you have no use or need for a monoculture in the first place, since a good organized clone culture will give you better results faster


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"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"
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OfflineTrippyLivin
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Re: Saving agar that's drying out [Re: c10h12n2o]
    #24262774 - 04/21/17 09:53 PM (7 years, 8 months ago)

Trust me I understand this is a long process. I estimated about a year time frame I was just curious where to start but I'll clone onto agar and do agar transfers. What are good nutrients to add? Could I do a LC from dunking water in grain jars if done sterile in front of a flow hood


--------------------
Happy trippin!

-TrippyLivin

Check out my indoor B+ and Golden Teacher Indoor Bulk Grow!
Adding AA+ Albino Cubensis,  Corumba Brazil Cubensis, Mazatapec Cubensis, and Texas PE6 Cubensis

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Re: Saving agar that's drying out [Re: TrippyLivin]
    #24262795 - 04/21/17 10:02 PM (7 years, 8 months ago)

none... just use standard MEA, anything you add is going to cause problems and not improve anything... MEA is good enough for labs, its plenty good for us

also, what you said about getting a monoculture from invitro stuff makes it seem like you dont know that those two things arent related. might want to do some reading up on the monocultures vs MS, or just realize that its irrelevant to the goal of growing lots of shrooms and stick to clone cultures :smile:

you could do an grain water LC, but its not a good idea... LC in general should be left to intermediate and up growers, they can cause all kinds of problems when you are getting started.

go spores > agar > liquid inoculant (not LC) > grain > CVG monotub > clone on agar > grain > CVG monotub w/ clone culture


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

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: Saving agar that's drying out [Re: c10h12n2o]
    #24263300 - 04/22/17 03:08 AM (7 years, 8 months ago)

Strain isolation is a lot of work but not actually as much as you would think, I mostly clone but I like agar work so I also isolate strains

8-10 transfers until there is really obvious sectoring usually

Let's say you take transfers from 10 sectors to new plates

Some of them might be isolates, usually IME I need another transfer or two until it stops sectoring.

Then each of those 10 cultures need to be tested. That's the long part IMO.

Uses a few sleeves of plates though!


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Re: Saving agar that's drying out [Re: mushpunx]
    #24263404 - 04/22/17 05:38 AM (7 years, 8 months ago)

Straight MEA or grain water if you saved some from making your jars. I used oat grain water last plates I poured and it worked great. 200 ml grain water, 300 ml distilled water, 10 grams agar agar. PCed 30 min @ 20 psi


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Re: Saving agar that's drying out [Re: CapnZ]
    #24263412 - 04/22/17 05:56 AM (7 years, 8 months ago)

I've never used grain water in agar  , I rotate between MEA and PDA.
Gunna save some grain water though give it a try- do you replace all the water or with GW or just some?


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Re: Saving agar that's drying out [Re: mushpunx]
    #24263461 - 04/22/17 07:10 AM (7 years, 8 months ago)

Just some. It's a 2/3 ratio. For 500 ml total I use 200 ml of grain water and 300 ml distilled water. I'm still experimenting too. I did one batch with just that, one batch like that with 1.5 grams of LME added. Both seem to be working fine.



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Invisiblec10h12n2o
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Re: Saving agar that's drying out [Re: mushpunx]
    #24264162 - 04/22/17 01:45 PM (7 years, 8 months ago)

Quote:

mushpunx said:
Strain isolation is a lot of work but not actually as much as you would think, I mostly clone but I like agar work so I also isolate strains

8-10 transfers until there is really obvious sectoring usually

Let's say you take transfers from 10 sectors to new plates

Some of them might be isolates, usually IME I need another transfer or two until it stops sectoring.

Then each of those 10 cultures need to be tested. That's the long part IMO.

Uses a few sleeves of plates though!




Of course everything you are saying is true in principle

But have you taken any of those supposed isolates and transferred then another 10 or 20 times?

If it was an isolate it would be the same even growth on every plate right?

IME , if you transfer those "isolated" sectors another 10 or 20 times they might look like an isolate for a minute but eventually they almost always form new sectors

Like I said, I cloned that monster (from a clone tub) and I'm still trying to isolate from it almost a year later

I think it's going to depend on how many strains are present in the initial sample and how compatible they are in forming groups

Also even though it is true in principle there could be a cube monoculture that didn't fruit or didn't fruit well, I have never seen a cubensis isolate which would not fruit, and I've heard people much more experience with isolates say the same. Probably says more about the hardiness of cubensis than anything else


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

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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