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Offlinekidcharlemagne
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Do you guys usually isolate clone mycelium to a monoculture before fruiting?
    #27290040 - 05/02/21 07:38 AM (3 years, 8 months ago)

Having a tough time getting an answer to this.  When you guys clone a fruit to agar, do you typically isolate the individual strains and fruit them all, fruit just the most aggressive, or will you fruit the combined strains?  Thanks.

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Re: Do you guys usually isolate clone mycelium to a monoculture before fruiting? [Re: kidcharlemagne]
    #27290107 - 05/02/21 08:28 AM (3 years, 8 months ago)

I dont follow what you are asking. Are you talking about sectoring? Cause if it is a clone it is the same strain across all the growth in the agar. I assume you are referring to P Cubes?

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Offlinekidcharlemagne
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Re: Do you guys usually isolate clone mycelium to a monoculture before fruiting? [Re: Sockadin]
    #27290171 - 05/02/21 09:21 AM (3 years, 8 months ago)

Quote:

Sockadin said:
I dont follow what you are asking. Are you talking about sectoring? Cause if it is a clone it is the same strain across all the growth in the agar. I assume you are referring to P Cubes?




Well the sectoring is indicative of multiple strains.  Clones usually have more than one strain for reasons I've never been able to get an answer for.  Yes cubes.

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Re: Do you guys usually isolate clone mycelium to a monoculture before fruiting? [Re: kidcharlemagne]
    #27290176 - 05/02/21 09:32 AM (3 years, 8 months ago)

You confused me with the term strain.

Yeah you can clone tissue from a mushroom and then isolate out sectors of growth to refine different genetic expressions in the fruit bodies but you do not know what you have until you grow it out. No where in this process is a "strain" isolated because it is all the same "Strain" P Cubes. It isn't like weed.

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Offlinekidcharlemagne
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Re: Do you guys usually isolate clone mycelium to a monoculture before fruiting? [Re: Sockadin]
    #27290212 - 05/02/21 10:13 AM (3 years, 8 months ago)

Quote:

Sockadin said:
You confused me with the term strain.

Yeah you can clone tissue from a mushroom and then isolate out sectors of growth to refine different genetic expressions in the fruit bodies but you do not know what you have until you grow it out. No where in this process is a "strain" isolated because it is all the same "Strain" P Cubes. It isn't like weed.




From my readings, any single dikaryotic hyphae is a "strain."  P. Cubensis is a species, and then you have "varieties" like B+, PE, etc. For reasons I can't find a answer to, a clone usually has more than one strain.  I'm assuming only one expresses the phenotype but I don't know. I trying to find out if people keep isolating sectors to get x number of monocultures and fruit each one to find the one that is contributing the genetics or if they fruit them as one polyculture.

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InvisibleStipe-n CapMDiscord
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Re: Do you guys usually isolate clone mycelium to a monoculture before fruiting? [Re: Sockadin]
    #27290225 - 05/02/21 10:25 AM (3 years, 8 months ago)

Strains and varieties are two different terms with different meanings.

A variety refers to the commercial name.

A strain is formed when compatible hyphae mate to form dikaryotic mycelium. Not all strains fruit, it would be difficult to produce a true isolated monoculture.


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Offlinekidcharlemagne
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Re: Do you guys usually isolate clone mycelium to a monoculture before fruiting? [Re: Stipe-n Cap]
    #27299801 - 05/08/21 07:41 AM (3 years, 8 months ago)

Gonna give this one a bump if you don't mind because I would think many of you would have an opinion.  Just trying to find out what you guys do once mycelium starts growing on agar from a fruit biopsy.  Grow it out as is, isolate sectors, etc?  Thanks.

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InvisibleStipe-n CapMDiscord
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Re: Do you guys usually isolate clone mycelium to a monoculture before fruiting? [Re: kidcharlemagne]
    #27299841 - 05/08/21 08:15 AM (3 years, 8 months ago)

When you put clone tissue to a plate it's wise to make at least one transfer to insure that it's clean, that's all you need to do.

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OfflineSub-Easy
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Re: Do you guys usually isolate clone mycelium to a monoculture before fruiting? [Re: Stipe-n Cap]
    #27299869 - 05/08/21 08:43 AM (3 years, 8 months ago)

Quote:

p9hu7 said:
When you put clone tissue to a plate it's wise to make at least one transfer to insure that it's clean, that's all you need to do.





I always just heard that you use the clone sample agar to inoculate grain. (After you know it's contamination free, of course.)

The process that I understand people use, would go something like this:
MS to agar, sector isolation, further sector isolation until you have many plates, grow out all plates, clone the best fruit and make LC or inoculate more dishes or grain with the dishes that produced the best mushrooms, and save the best agar dishes. Those dishes are your monoculture and master LC.

Is there more?

What am I missing?

Edited by Sub-Easy (05/08/21 09:01 AM)

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InvisibleStipe-n CapMDiscord
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Re: Do you guys usually isolate clone mycelium to a monoculture before fruiting? [Re: Sub-Easy]
    #27299877 - 05/08/21 08:48 AM (3 years, 8 months ago)

When you take a clone it's growing in open air and you're handling it with your dirty mits. You may contaminate the sample when taking the tissue from the fruit, but perhaps not. I've taken clean tissue culture's on many occasions but it's good practice to transfer away from the original clone plate to insure that you don't have any contamination on the plate.

You don't need to try to make a monoculture from that plate, doubtful that you could anyways. Even if you could get a monoculture it would defeat the purpose of taking a clone.

Use the clean plate in the same manner you would use any other clean plate.

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Re: Do you guys usually isolate clone mycelium to a monoculture before fruiting? [Re: Stipe-n Cap]
    #27299917 - 05/08/21 09:14 AM (3 years, 8 months ago)

Quote:

p9hu7 said:
When you take a clone it's growing in open air and you're handling it with your dirty mits. You may contaminate the sample when taking the tissue from the fruit, but perhaps not. I've taken clean tissue culture's on many occasions but it's good practice to transfer away from the original clone plate to insure that you don't have any contamination on the plate.

You don't need to try to make a monoculture from that plate, doubtful that you could anyways. Even if you could get a monoculture it would defeat the purpose of taking a clone.

Use the clean plate in the same manner you would use any other clean plate.




Yep, you are right.
I was using your quote as an example of how I understand it to work also.

Once you clone, that's about as far as iv hard of someone taking it.

Or the other way is MS/grain sectoring/genetic isolation.

Is there anything more to it that I hadn't heard of?

Excluding mushroom hybrids.


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InvisibleStipe-n CapMDiscord
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Re: Do you guys usually isolate clone mycelium to a monoculture before fruiting? [Re: Sub-Easy]
    #27299944 - 05/08/21 09:34 AM (3 years, 8 months ago)

When you transfer from an ms germ plate you're not attempting any significant genetic isolation other than isolation from contamination.

Any transfer you care to take with your scalpel will probably have 10⁴+ strains present in the sample. So genetic isolation is not that significant while making ordinary transfers. This is why your clone remains a clone even though you transferred from the original tissue sample plate.

That's pretty much all you need to know about making transfers for simple cultivation purposes.

I wouldn't worry about isolating strains and monoculture. What matters is the cleanliness of your culture and it's fruiting efficiency.

You're basically just cloning for Efficiency/potency and maybe selecting for phenotype when considering crosses to make.

Edited by Stipe-n Cap (05/08/21 09:44 AM)

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Offlinekidcharlemagne
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Re: Do you guys usually isolate clone mycelium to a monoculture before fruiting? [Re: Stipe-n Cap]
    #27302543 - 05/10/21 07:00 AM (3 years, 8 months ago)

Quote:

p9hu7 said:
When you transfer from an ms germ plate you're not attempting any significant genetic isolation other than isolation from contamination.

Any transfer you care to take with your scalpel will probably have 10⁴+ strains present in the sample. So genetic isolation is not that significant while making ordinary transfers. This is why your clone remains a clone even though you transferred from the original tissue sample plate.

That's pretty much all you need to know about making transfers for simple cultivation purposes.

I wouldn't worry about isolating strains and monoculture. What matters is the cleanliness of your culture and it's fruiting efficiency.

You're basically just cloning for Efficiency/potency and maybe selecting for phenotype when considering crosses to make.




But if you have 10 strains on that plate and fruit them all collectively, wouldn't any fruit have only 1/10 the chance of exhibiting the pheno of your original clone?

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InvisibleStipe-n CapMDiscord
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Re: Do you guys usually isolate clone mycelium to a monoculture before fruiting? [Re: kidcharlemagne]
    #27302548 - 05/10/21 07:04 AM (3 years, 8 months ago)

I don't really follow, but you're not going to have 10, I said 10⁴+ (10,000+). There are many thousands of individual strains all overlapping and winding around each other in a visable rhizomorphic rope of mycelium.

Individual hyphae are microscopic, the fact that you can see them as a "rope" like structure on your agar is an indication of just how many are potentially present not only on your plate, but in each transfer, no matter how small you care to cut it.

"Hyphae have an average diameter of 4–6" µm.
Source: Google.

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