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Invisiblegoldieman
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Registered: 08/15/07
Posts: 296
Loc: midwest
Mushroom tissue to agar.. Is this an isolate??
    #16844534 - 09/14/12 12:55 AM (12 years, 4 months ago)

I'm starting a new grow. I "knocked up" 9 petri dishes, 3 of each strain, with spore syringes. I plan on taking a piece of agar to make LC's of each strain, use the LC's to inoculate grain jars, then grow out a batch of each strain. From there I plan to take tissue samples of fruits from each strain and put those pieces on agar. My question is: Are the tissue samples going to grow out isolates, or do I have to sector further from there??

Also, how long should it take for spores to germinate on agar? How much growth should I wait for before I take a sample from agar to make LC"s?? AM I going about this the right way???

A few words of encouragement would be nice! LOL. Thanx to all who reply with valid info!!

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OfflineIamYou
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Registered: 10/11/11
Posts: 704
Last seen: 8 years, 4 months
Re: Mushroom tissue to agar.. Is this an isolate?? [Re: goldieman]
    #16845155 - 09/14/12 07:41 AM (12 years, 4 months ago)

from what ive heard, not that understand why, but i think you will still need to isolate and grow it out on other plates.  surely someone with some actual experience  with this will chime in soon.


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“Life lived in the absence of the psychedelic experience is life trivialized, life denied, life enslaved to the ego.”
- You


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Offlinemindraped
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Re: Mushroom tissue to agar.. Is this an isolate?? [Re: IamYou]
    #16845228 - 09/14/12 08:11 AM (12 years, 4 months ago)

A single fruit can be made up of more than one substrain.

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OfflineMush Freak
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Registered: 12/29/11
Posts: 154
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Re: Mushroom tissue to agar.. Is this an isolate?? [Re: mindraped]
    #16845263 - 09/14/12 08:28 AM (12 years, 4 months ago)

Im curious why you started on agar and then to LC?

Since you wont really know what your growing until you isolate from a fruit body youve already grown, starting from an LC may be a faster way to go. After you have a good specimen to use then place tissue on agar and isolate.

This way you know your sample will have the traits yiur looking for. I.e. speed of growth, size, potency, sporalation...

Goodluck and let use know how this turns out.


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:mushroom2::mushroom2:Mono-tubs are LIFE:mushroom2::mushroom2:


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OfflinePussyFart
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Registered: 04/08/12
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Re: Mushroom tissue to agar.. Is this an isolate?? [Re: mindraped] * 1
    #16845268 - 09/14/12 08:29 AM (12 years, 4 months ago)

An isolate is a mycellium that grows out evenly.(No Sectoring)

A single fruit body can consist of upwars of 20 different "substrains" which can each be isolated on agar.

When u place a tissue sample on agar, chances are there will be multiple "sectors" growing from it.

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OfflinePussyFart
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Re: Mushroom tissue to agar.. Is this an isolate?? [Re: Mush Freak] * 1
    #16845270 - 09/14/12 08:31 AM (12 years, 4 months ago)

Quote:

Mush Freak said:
Im curious why you started on agar and then to LC?

Since you wont really know what your growing until you isolate from a fruit body youve already grown, starting from an LC may be a faster way to go.




Starting an LC from spores is a crapshoot because the cap from which the spores came was fruited in the open air.

No spore print/syringe is ever 100% clean, this is why we start spores on agar, so we can make sure its clean.

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OfflineMush Freak
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Re: Mushroom tissue to agar.. Is this an isolate?? [Re: PussyFart]
    #16845510 - 09/14/12 09:46 AM (12 years, 4 months ago)

[Quote]Notahacker420 said:
Quote:

Mush Freak said:
Im curious why you started on agar and then to LC?

Since you wont really know what your growing until you isolate from a fruit body youve already grown, starting from an LC may be a faster way to go.




Starting an LC from spores is a crapshoot because the cap from which the spores came was fruited in the open air.

No spore print/syringe is ever 100% clean, this is why we start spores on agar, so we can make sure its clean.



Good point!



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:mushroom2::mushroom2:Mono-tubs are LIFE:mushroom2::mushroom2:


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OfflineGoldenArrow
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Re: Mushroom tissue to agar.. Is this an isolate?? [Re: Mush Freak]
    #16845532 - 09/14/12 09:52 AM (12 years, 4 months ago)

I wouldn't go to LC though, go to grain then GLC or G2G

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OfflinePussyFart
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Re: Mushroom tissue to agar.. Is this an isolate?? [Re: GoldenArrow]
    #16845563 - 09/14/12 09:58 AM (12 years, 4 months ago)

Quote:

GoldenArrow said:
I wouldn't go to LC though, go to grain then GLC or G2G



All I do is agar and G2G, I do not use LCs, no reason too.

I do spores to pint of grains, then G2G that to 3 quarts, then G2G those to 30. And bam, 6 or 7 mono tubs of spawn in a month to a month and a half.

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Offlinesgfcchamber
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Re: Mushroom tissue to agar.. Is this an isolate?? [Re: goldieman] * 1
    #16845900 - 09/14/12 11:11 AM (12 years, 4 months ago)

Quote:

goldieman said:
I'm starting a new grow. I "knocked up" 9 petri dishes, 3 of each strain, with spore syringes. I plan on taking a piece of agar to make LC's of each strain, use the LC's to inoculate grain jars, then grow out a batch of each strain. From there I plan to take tissue samples of fruits from each strain and put those pieces on agar. My question is: Are the tissue samples going to grow out isolates, or do I have to sector further from there??

Also, how long should it take for spores to germinate on agar? How much growth should I wait for before I take a sample from agar to make LC"s?? AM I going about this the right way???

A few words of encouragement would be nice! LOL. Thanx to all who reply with valid info!!




The first thing I would recommend, is to skip LC altogether.  They are notoriously sketchy and even when innoculating with an agar wedge, you still have to worry about contamination of the LC...and you don't find out it's contaminated until it's too late.

If you've got a clean culture on agar, I'd go straight to a grain jar, then G2G to expand the mycelium.  Now, to answer your other questions:

Quote:

My question is: Are the tissue samples going to grow out isolates, or do I have to sector further from there??




While a cloned fruit CAN grow out as an isolate, it's very likely that it won't and that you'll need to do a couple of transfers to get to one.

Quote:

Also, how long should it take for spores to germinate on agar?




I typically see growth from spores on agar in about 4 to 5 days.  It can take more or less though, that's just my "average".  If you're innoculating the dishes with a spore syringe, be absolutely sure to use no more than a single very small drop, and then also make sure that "drop" doesn't "run" across the plate.  Just make sure it sits right where you set it.  If there is any bacteria present, it's going to flourish in the water, so you want to make sure you don't go spreading it all over the place.

Quote:

How much growth should I wait for before I take a sample from agar to make LC"s??




When starting from spores, you should really wait until they germinate on the plate, then make a transfer to another plate as soon as you can.  This ensures that you've got a clean culture.  Spore syringes are almost never (if ever) 100% "clean" and there could be a "hidden" contaminate underneath your healthy looking mycelium on "plate 0", so you should always transfer from the germination plate to another plate for more growth so you can absolutely guarantee you've got a clean culture.

As I said, I would skip the LC altogether, and go straight from agar to a grain jar.  I let my cultures grow to about an inch or so within the edge of the plate, and then cut the plate into 4 pieces, and then cut one of those four pieces into 3 pieces, and inoculate the jar with 1 to all 3 of those pieces.

Now, for some pretty pictures showing what I got when I cloned a fruit to agar, to demonstrate that no, you don't necessarily get a monoculture when cloning a fruit (here is the thread these pics were originally posted on, just in case you want to see ALL the pictures along with info: This is What Happens When You Take Tissue from an Adult Fruit and Put it on Agar (with pics) )


CDB1-1:


CDB1-1 (taken from bottom of dish):


CDB1-2:


CDB1-2 (taken from bottom of dish):


CDB2-1:


CDB2-1 (taken from bottom of dish):


CDB2-2:


CDB2-3:


SDB-3:


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