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OfflineClc420
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Confusion with cloning and isolating
    #26617079 - 04/21/20 08:30 AM (3 years, 9 months ago)

I understand what cloning and isolating is. I took my first clone today on agar. What confusing me is I see some people say they prefer growing clones than isolates. When the tissue culture grows out eventually it will grow out in sectors. If you wasnt going to isolate it surely you would end up taking a sector when transferring to a new dish ?

So if you just wanted to keep the original clone culture how would you do that without isolating down the road ?


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OfflineA.k.aM
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Re: Confusion with cloning and isolating [Re: Clc420]
    #26617195 - 04/21/20 09:19 AM (3 years, 9 months ago)

I think with isolate they mean it’s down to one set of genetics. It’s super confusing with isolate and monoculture being used different ways.

If you have a master plate of a good clone, my strategy is to take transfers close to the center so there’s more genetics in it.

I recently started putting a wedge down and letting it grow for a week, then cutting the wedge out and putting it in a new plate. That way it’s the exact same culture/genetics on every plate.

Idk how long you can do that for before it gets tired though.


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Edited by A.k.a (04/21/20 09:20 AM)


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OfflineMadoxx
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Re: Confusion with cloning and isolating [Re: Clc420]
    #26617362 - 04/21/20 10:19 AM (3 years, 9 months ago)

As A.k.a said if you want to avoid isolating a 'clone' when transferring between plates you should try and take tissue from the center of the plate. This is because you have to imagine mycelium as a load of interlinked hyphae in which every one is susceptible to have a different genetics from the one next to it. This is specially in grows from MS where there are a number of different genetics present, equal to the genetic diversity found in the spores from which you inoculated from.

When you 'clone' you are taking a piece of a 'mostly' similar genetic sample from a cap for example, and the other genetics which ride along in that fruit. If after 'cloning' on agar a few times the mycelium shows signs of being tired just go ahead and fruit a sample to renew the vigor of the culture. You will also get a ''pure'' sample of your ''clone''.
If you have read until here i want to stress the " ' ". In mycology clones are only so much a % equal (what we understand by clone in animals for example) due to varying proportions of genetics found in non-monoculture clones.'Pure' meaning you are going to get a more or less identical proportion of genetics to the ones found in your clone the last time you took a sample, because you are not selecting for any special genetics as you mention can happen on an agar plate eventually.


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Offlinepoisoned
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Re: Confusion with cloning and isolating [Re: Madoxx]
    #26617392 - 04/21/20 10:30 AM (3 years, 9 months ago)

To keep "fresh cultures", you have to paralelyze your agar work. Create a master plate, create slants from it and then create slave plates. Once you run out of your master plate, go to your fridge and you'll have young culture in that slant.


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OfflineClc420
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Re: Confusion with cloning and isolating [Re: poisoned]
    #26617456 - 04/21/20 10:56 AM (3 years, 9 months ago)

Quote:

poisoned said:
To keep "fresh cultures", you have to paralelyze your agar work. Create a master plate, create slants from it and then create slave plates. Once you run out of your master plate, go to your fridge and you'll have young culture in that slant.





That's what I thought could be a possibility is ti just make a slant with the clone culture and put it in the fridge.

With cubensis being a sub tropical species have you had any problem with master plates going into cold storage? Rr said he had lost p cubensis cultures from temps 35 and below. He recommends 50f for long term storage of cubensis cultures.


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Offlinepoisoned
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Re: Confusion with cloning and isolating [Re: Clc420]
    #26617528 - 04/21/20 11:28 AM (3 years, 9 months ago)

35 is almost freezing, your fridge shouldn't be that cold. I've only been doing any culture work for a bit more than a year. I did make slants, but I managed to contaminate them later on and didn't have any to continue storage with them. I did have masters in fridge for a few months and they worked.


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OfflineA.k.aM
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Re: Confusion with cloning and isolating [Re: poisoned]
    #26617807 - 04/21/20 01:27 PM (3 years, 9 months ago)

I can vouch for around 50.

I left town for the holidays for 10 days and when I came back my thermometer said the temp swing was 44-56 and it was like my jars and plates were frozen in time. I was all excited to come back to colonized stuff lol.


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OfflineClc420
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Re: Confusion with cloning and isolating [Re: A.k.a]
    #26617990 - 04/21/20 02:38 PM (3 years, 9 months ago)

Quote:

A.k.a said:
I can vouch for around 50.

I left town for the holidays for 10 days and when I came back my thermometer said the temp swing was 44-56 and it was like my jars and plates were frozen in time. I was all excited to come back to colonized stuff lol.





So myc grew at those temperatures? I want to keep slants and plates at 50 to halt growth for long term storage.


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OfflineA.k.aM
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Re: Confusion with cloning and isolating [Re: Clc420]
    #26618076 - 04/21/20 03:22 PM (3 years, 9 months ago)

Barely.

Tubs I had already spawned went like normal but plates and jars hardly moved.


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