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InvisibleMUSH HEAD420
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Re: Too slow for agar to grain? [Re: SpitballJedi]
    #19175288 - 11/22/13 03:30 PM (10 years, 3 months ago)

:whoah:


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Offlineflipsidetrue
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Re: Too slow for agar to grain? [Re: MUSH HEAD420]
    #19175357 - 11/22/13 03:47 PM (10 years, 3 months ago)

i'll be following this thread because i used some colonized grain from a print i made that was derived from a MS syringe to agar to isolate, never thought about how it could affect the growth though when doing multiple g2g transfer to keep the strain going, is there a certain amount of times that you can actually do this before the strain stops producing?


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Edited by flipsidetrue (11/22/13 03:55 PM)


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InvisibleMUSH HEAD420
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Re: Too slow for agar to grain? [Re: flipsidetrue]
    #19175394 - 11/22/13 03:58 PM (10 years, 3 months ago)



I am guessing to get a good isolate you must use as few spores as possible, that is the key.

I didn't know that genes from the multispore fruits will only isolate down to multiple genes from the multispore.

I figured if you got an albino from a multispore and cloned it, you may have a higher rate of albinos.


Edited by MUSH HEAD420 (11/22/13 03:59 PM)


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InvisibleSpitballJedi
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Re: Too slow for agar to grain? [Re: flipsidetrue]
    #19175448 - 11/22/13 04:09 PM (10 years, 3 months ago)

Quote:

flipsidetrue said:is there a certain amount of times that you can actually do this before the strain stops producing?




It depends on genetics.


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InvisibleMUSH HEAD420
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Re: Too slow for agar to grain? [Re: SpitballJedi]
    #19176067 - 11/22/13 06:56 PM (10 years, 3 months ago)

So I could take a wedge of colonized agar from a MS clone, make a liquid culture with it, then inoculate grain to spawn to bulk?

Would the resulting fruits be similar to the clone?
Quote:

cronicr said:
Yep




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Invisibleblindingleaf
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Re: Too slow for agar to grain? [Re: MUSH HEAD420]
    #19176115 - 11/22/13 07:08 PM (10 years, 3 months ago)

in that first pic, it looks like the myc is just the tip of the wedge maybe it just wasn't enough myc to take off super fast?  usually when i do agar transfer, I don't shake, but I slowly wobble the jar back and forth till the wedge is not on the surface, but not deep at all, so grains are totally covering it.  my apt is also at 65 so I have exact problem with slow growth, but  once the agar myc has traveled to 10-15 grains, Ill shake.  nothing looks wrong, I wouldn't toss yet, but nothing wrong with starting more jars! just use bigger agar wedge next time


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InvisibleSpitballJedi
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Re: Too slow for agar to grain? [Re: MUSH HEAD420]
    #19176277 - 11/22/13 07:47 PM (10 years, 3 months ago)

Quote:

MUSH HEAD420 said:
So I could take a wedge of colonized agar from a MS clone, make a liquid culture with it, then inoculate grain to spawn to bulk?

Would the resulting fruits be similar to the clone?
Quote:

cronicr said:
Yep







Yep.


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OfflinePussyFart
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Re: Too slow for agar to grain? [Re: SpitballJedi]
    #19176401 - 11/22/13 08:23 PM (10 years, 3 months ago)

Quote:

SpitballJedi said:
A fruit from a MS grow has multiple genetics.

A clone from a MS grow will not likely be an isolate.

An isolate made from a clone of a MS fruit can result in failure because the multiple genetics are no longer working in concert.



:whathesaid:


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OfflinePsilicon
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Re: Too slow for agar to grain? [Re: SpitballJedi]
    #19176434 - 11/22/13 08:33 PM (10 years, 3 months ago)

Quote:

SpitballJedi said:
Due to genetics, some will die out sooner than others. They can only go through so many cell divisions.





Your theory is it's a telomerase problem, as with senescence in humans?


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InvisibleSpitballJedi
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Re: Too slow for agar to grain? [Re: Psilicon]
    #19177746 - 11/23/13 05:09 AM (10 years, 3 months ago)

It was initially. but new info lead me another direction.


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Edited by SpitballJedi (11/23/13 05:37 AM)


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InvisiblebodhisattaMDiscordReddit
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Re: Too slow for agar to grain? [Re: SpitballJedi]
    #19177873 - 11/23/13 07:11 AM (10 years, 3 months ago)

Quote:

SpitballJedi said:
A fruit from a MS grow has multiple genetics.

A clone from a MS grow will not likely be an isolate.

An isolate made from a clone of a MS fruit can result in failure because the multiple genetics are no longer working in concert.




That's what I really needed to know.

I wasn't worried about achieving rhizo groth it was rhizo right from the fruit body. I just wanted to work down to an isolate. But I have heard to use MS fruits to work to get an isolate since you're at least working with strains that are known to fruit rather than starting from spores.

I knew I would still have to do more than 1 transfer to get to an isolate even with an MS fruit and that's what I was doing. Perhaps the mycelium has ran into a wall since I removed it from its friend strains.

and yea my grains are on the dry side.

So if I want an isolate start with spores not a MS fruit? Especially since the MS fruit was from a bulk grow which has already undergone a assload of mycelial expansion.


Edited by Trusted cuItivator (11/23/13 07:14 AM)


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Invisibleblindingleaf
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Re: Too slow for agar to grain? [Re: bodhisatta]
    #19177939 - 11/23/13 07:43 AM (10 years, 3 months ago)

I dunno if this helps, but I just isolate, whether from fruit or spore, then noc up a pint jar of WBS and grow it out with coir/verm (because thats what I'm going to do anyway when I go bigger) to see how it performs.  I use these small buckets that are mock mono's, so I have several going at the same time.  1 pint spawn to 1 pint pasteurized sub.  some do well some do ok some do awesome.  The awesome ones, I keep the iso. 

My thought process (correct me if I'm wrong) is if I'm going to spawn to coir/verm using WBS then I wanna make sure that the iso likes WBS and coir/verm (as well as if it fruits, potency, yield, etc).  I think some ppl do cakes for this process, but since I primarily use WBS, then coir/verm I wanna make sure that iso can perform well utilizing those specific nutrient mediums.  I've read posts where some myc does well on MEA but not so well on PDYA, etc, so I guess I'm using the same process, but with spawn nutrient than with agar nutrients.  Pastywhite mentions this at times when discussing agar nutrients/agar work
http://www.shroomery.org/forums/showflat.php/Number/19149708
(sorry pasty, but this was very informative to me, so Im throwing it out there)


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Invisiblegrainbrain
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Re: Too slow for agar to grain? [Re: blindingleaf]
    #19178206 - 11/23/13 09:44 AM (10 years, 3 months ago)

Quote:

blindingleaf said:
I slowly wobble the jar back and forth till the wedge is not on the surface, but not deep at all, so grains are totally covering it.




This is what works for me as well.  Dropping a wedge on top of grains has rarely performed as well as rolling the wedge down into the grains against the glass.


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Re: Too slow for agar to grain? [Re: grainbrain]
    #19178621 - 11/23/13 11:46 AM (10 years, 3 months ago)

Yes you can get an iso from spores but I take a few as they need to be tested, when working with a clone you gotta watch for sectoring, I still make a quick transfer to make sure its clean but I take a generous piece, on the other hand even when they do sector the myc still spreads so I'm not sure that's your issue here


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InvisibleSpitballJedi
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Re: Too slow for agar to grain? [Re: bodhisatta]
    #19178732 - 11/23/13 12:18 PM (10 years, 3 months ago)

Quote:

SpitballJedi said:
A fruit from a MS grow has multiple genetics.

A clone from a MS grow will not likely be an isolate.

An isolate made from a clone of a MS fruit can (but not always) result in failure because the multiple genetics are no longer working in concert.




I clarified that a little.


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Edited by SpitballJedi (11/23/13 12:18 PM)


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InvisiblebodhisattaMDiscordReddit
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Re: Too slow for agar to grain? [Re: SpitballJedi]
    #19179703 - 11/23/13 05:05 PM (10 years, 3 months ago)

Quote:

blindingleaf said:
I dunno if this helps, but I just isolate, whether from fruit or spore, then noc up a pint jar of WBS and grow it out with coir/verm (because thats what I'm going to do anyway when I go bigger) to see how it performs.  I use these small buckets that are mock mono's, so I have several going at the same time.  1 pint spawn to 1 pint pasteurized sub.  some do well some do ok some do awesome.  The awesome ones, I keep the iso. 

My thought process (correct me if I'm wrong) is if I'm going to spawn to coir/verm using WBS then I wanna make sure that the iso likes WBS and coir/verm (as well as if it fruits, potency, yield, etc).  I think some ppl do cakes for this process, but since I primarily use WBS, then coir/verm I wanna make sure that iso can perform well utilizing those specific nutrient mediums.  I've read posts where some myc does well on MEA but not so well on PDYA, etc, so I guess I'm using the same process, but with spawn nutrient than with agar nutrients.  Pastywhite mentions this at times when discussing agar nutrients/agar work
http://www.shroomery.org/forums/showflat.php/Number/19149708
(sorry pasty, but this was very informative to me, so Im throwing it out there)




this is pretty much what I do but I use rye instead. The thing it's now so far away from when it was a spore that is that the reason why I'm seeing this poor performance?


Quote:

grainbrain said:
Quote:

blindingleaf said:
I slowly wobble the jar back and forth till the wedge is not on the surface, but not deep at all, so grains are totally covering it.




This is what works for me as well.  Dropping a wedge on top of grains has rarely performed as well as rolling the wedge down into the grains against the glass.




I suggested other people do this as well and then RR schooled me about it. You should drop a wedge onto the grain and let it start from there rather than roll it right away.

Quote:

RogerRabbit said:
Those look fine, but you would have been better off not to shake as directed above.

The reason is that when you cut off a wedge, you cut through the growing, leading edge of the mycelium.  It's best to let this recover without further damage, so that the mycelium crawls off the wedge and into the grains below.  It makes little difference if the wedge is upside down or right side up because it's not going to recover and grow from the middle, but from the leading edges.

Once the mycelium has crawled into a few of the grains below, shake to distribute.  Shaking early with a mycelium wedge slows things down because you only get recovery later from the wedge itself, not from all the mycelium which falls off when you shake, but doesn't recover and grow.  You must then wait for what's left on the agar wedge to recover and grow again before the mycelium can leap off, setting things back a few days.

The other advantage to not shaking right away is that you can see the agar wedge on top so that if green mold or another contaminant develops, you see it before you shake so you can discard that jar.
RR





Quote:

cronicr said:
Yes you can get an iso from spores but I take a few as they need to be tested, when working with a clone you gotta watch for sectoring, I still make a quick transfer to make sure its clean but I take a generous piece, on the other hand even when they do sector the myc still spreads so I'm not sure that's your issue here




I haven't seen sectoring happen yet and I'm 3 dishes into a MS fruit clone. Perhaps it was already an isolate from the 1st transfer because the plates did look sort of like some peoples isolate strain plates?

Quote:

SpitballJedi said:
Quote:

SpitballJedi said:
A fruit from a MS grow has multiple genetics.

A clone from a MS grow will not likely be an isolate.

An isolate made from a clone of a MS fruit can (but not always) result in failure because the multiple genetics are no longer working in concert.




I clarified that a little.




I guess I wonder if I'm in the "failure" part of that or something else.


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Invisibleblindingleaf
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Re: Too slow for agar to grain? [Re: bodhisatta]
    #19179718 - 11/23/13 05:10 PM (10 years, 3 months ago)

I guess i just see faster growth when the wedge is covered in grain, I don't wobble it hard enough to damage, just let the grains gently come on top of the wedge.
as far as being so far from spore to effect performance, I dunno.  Actually, I wasn't even aware that trying to isolate something from a fruit clone could end up being detrimental until I read this thread.  I've been isolating from clones for last 2 weeks, so I guess all that work might not pay off.


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Re: Too slow for agar to grain? [Re: blindingleaf]
    #19179732 - 11/23/13 05:14 PM (10 years, 3 months ago)

Ok so my revised question list is

1. Clone tissue from a MS grow is sub optimal to obtain an isolate from?
if you use clone tissue from MS the best practice to use it for is take one transfer to a new dish and use that whole next dish to inoculate grain.

2. You think for the reasons in question no1 I'm seeing poor performance from these agar wedges?

3. If you want an isolate you should start from a very small(as small as possible) amount of spores onto agar. Then try out the isolates after they're obtained to see their fruiting characteristics?


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InvisibleMUSH HEAD420
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Re: Too slow for agar to grain? [Re: bodhisatta]
    #19180167 - 11/23/13 07:16 PM (10 years, 3 months ago)

Just think of the clone as the isolate, it's entire genetic makeup is what you are after. If you clone a huge fruit you do not want to isolate it any because the genetic makeup of the clone sample is what you are after.

Now if you start from spores in agar, you use as few spores as possible to get the least amount of germination. Then sector out the best looking sample and that is your isolate.

I'm sure you can use the agar with the tissue sample, as long as it does not show contamination. Mycelium is contam resistant and when starting from a clone it goes quick.



Edited by MUSH HEAD420 (11/23/13 07:28 PM)


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InvisiblebodhisattaMDiscordReddit
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Re: Too slow for agar to grain? [Re: MUSH HEAD420]
    #19180183 - 11/23/13 07:22 PM (10 years, 3 months ago)

Looks like I have some work to do. To the SAB, man the PC!


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