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Offlineiperuranio
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Registered: 10/30/12
Posts: 6
Last seen: 11 years, 2 months
LC progressive diluition
    #17375387 - 12/11/12 04:23 PM (11 years, 5 months ago)

Hi guys,
i was wondering about a procedure so i tought about asking to you.
Imagine we have, for example, 5 vial of sterilized liquid substrate of 9 ml and a MS Liquid culture ready to use.
I pick 1 ml from the LC and inject it in the first vial, wait for growth, shake, than pick 1 ml from that one and inject in the next one ecc.
Does the micelium that grows in the last one will be more "pure"?, i mean, do you think there will be less different strains?

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OfflineTmethylM
Smear in the shale
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Registered: 07/16/12
Posts: 16,431
Loc: Florida
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Trusted Cultivator
Re: LC progressive diluition [Re: iperuranio]
    #17375458 - 12/11/12 04:36 PM (11 years, 5 months ago)

So you're talking about isolating strains through use of liquid cultures.
It's partially possible, you can get a 'close to monoculture' vial but there is no way to know, and by the second or third culture you will surely have ruined the vials with contamination. This is why Agar is so beautiful, you can SEE your sectoring and isolate the better strains, and you can SEE contamination and isolate away from it and clean up your cultures yourself.

This is kind of possible, but in n o way practical or sane. :tongue:
I'd give this a 5-10% chance of being remotely successful. (With great sterile procedures and experience)


--------------------
¯\_(ツ)_/¯

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Offlineiperuranio
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Registered: 10/30/12
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Re: LC progressive diluition [Re: Tmethyl]
    #17378861 - 12/12/12 09:15 AM (11 years, 5 months ago)

Thank you, very clear answer. It was just a theoretical question.

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OfflineTmethylM
Smear in the shale
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Registered: 07/16/12
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Re: LC progressive diluition [Re: iperuranio]
    #17378973 - 12/12/12 09:42 AM (11 years, 5 months ago)

Wasn't trying to appear sarcastic or disrespectful.
Its a valid question and it could have some practical applications if you had a sterile lab and proper equipment to work with.
But agar is so simple, effective, and safe in comparison. The major downfall of any LC is the contamination, they contam very easy and its impossible to know if they are contaminated when you inoculate. Huge risk especially for large grows or commercial operations.


--------------------
¯\_(ツ)_/¯

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Offlineskippydude
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Registered: 09/08/12
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Re: LC progressive diluition [Re: iperuranio]
    #17378988 - 12/12/12 09:46 AM (11 years, 5 months ago)

Quote:

iperuranio said:
Hi guys,
i was wondering about a procedure so i tought about asking to you.
Imagine we have, for example, 5 vial of sterilized liquid substrate of 9 ml and a MS Liquid culture ready to use.
I pick 1 ml from the LC and inject it in the first vial, wait for growth, shake, than pick 1 ml from that one and inject in the next one ecc.
Does the micelium that grows in the last one will be more "pure"?, i mean, do you think there will be less different strains?




This is the recipe for senescence, not strain isolation!

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OfflineJeff
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Registered: 10/06/12
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Re: LC progressive diluition [Re: skippydude]
    #17382301 - 12/12/12 08:05 PM (11 years, 5 months ago)

Quote:

skippydude said:
Quote:

iperuranio said:
Hi guys,
i was wondering about a procedure so i tought about asking to you.
Imagine we have, for example, 5 vial of sterilized liquid substrate of 9 ml and a MS Liquid culture ready to use.
I pick 1 ml from the LC and inject it in the first vial, wait for growth, shake, than pick 1 ml from that one and inject in the next one ecc.
Does the micelium that grows in the last one will be more "pure"?, i mean, do you think there will be less different strains?




This is the recipe for senescence, not strain isolation!




Not likely skippy.  I have slants that were ~P-45 from Aloha that are still very aggressive.  Small transfers to small volumes will not lead to senescence, at least not for the 5 he has listed above.


--------------------
Myco-tek

Edited by Jeff (12/12/12 08:06 PM)

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Offlineskippydude
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Registered: 09/08/12
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Re: LC progressive diluition [Re: Jeff]
    #17382748 - 12/12/12 09:14 PM (11 years, 5 months ago)

This is still not the way to isolate a strain.
You could wear a culture out trying this way.
That is all I am saying.

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