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Offlinesemaphore
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Pan.Cyan jam agar plate, when to stop isolating.
    #14417913 - 05/08/11 04:26 AM (10 years, 8 days ago)

As the title says over the last 2 and a bit weeks i have been growing pans in agar to get a nice growth going, i have isolated my 2 dishes 3 times. So basically the dishes are in there 3rd isolation, one or two of them have contams, the other has cobweb mold. The dishes i cut from are still growing strong. What am i looking for with regards to this species, should i just cut a piece of it off now and do an LC or transfer a slice to WBS ?


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Invisiblecc2
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Registered: 05/15/10
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Re: Pan.Cyan jam agar plate, when to stop isolating. [Re: semaphore]
    #14418071 - 05/08/11 05:15 AM (10 years, 8 days ago)

Quote:

semaphore said:
one or two of them have contams, the other has cobweb mold. The dishes i cut from are still growing strong. What am i looking for with regards to this species




if at 3rd isolation you're still getting contams. either isolate more or improve your sterile technique.

looks silly inoculating to WBS or worse making a LC out of something which isn't sure to contain only what you're trying to grow.


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Offlinesemaphore
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Re: Pan.Cyan jam agar plate, when to stop isolating. [Re: cc2]
    #14418080 - 05/08/11 05:21 AM (10 years, 8 days ago)

Quote:

cc2 said:
Quote:

semaphore said:
one or two of them have contams, the other has cobweb mold. The dishes i cut from are still growing strong. What am i looking for with regards to this species




if at 3rd isolation you're still getting contams. either isolate more or improve your sterile technique.

looks silly inoculating to WBS or worse making a LC out of something which isn't sure to contain only what you're trying to grow.





Okay you didnt read it properly 2 out of 10 dishes have contams the rest are growing healthy (granted i never mentioned my total amount of 3rd isolation dishes), obviously i am not going to isolate or innoc from a contaminated dish :rolleyes: And its not contams of the mushroom culture itself its on the very outer edge some bacteria must have dropped onto the dish while i was working it, i suspect it may be the way i moved my hands over.


Edited by semaphore (05/08/11 05:23 AM)


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Invisiblecc2
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Re: Pan.Cyan jam agar plate, when to stop isolating. [Re: semaphore]
    #14418099 - 05/08/11 05:32 AM (10 years, 8 days ago)

Quote:

semaphore said:
Okay you didnt read it properly




of course, you couldn't be the one not explaining properly :rolleyes:

Quote:

semaphore said:
obviously i am not going to isolate or innoc from a contaminated dish :rolleyes: And its not contams of the mushroom culture itself its on the very outer edge some bacteria must have dropped onto the dish while i was working it




AFAIK contams. don't have to necessairly be over your mycelium for calling a plate contaminated. in fact most contams. unless it's cobweb will grow around your myc, not over it.

now let's also say bacteria are one of the easiest contams. to isolate healthy myc from, but it you had mold on the very outer edge it won't be so straightforward to inoculate with that.

if you've got some dishes that are contam-less, what's stopping you from inoculating then?


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Invisibleslapphappypill
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Re: Pan.Cyan jam agar plate, when to stop isolating. [Re: cc2]
    #14418104 - 05/08/11 05:34 AM (10 years, 8 days ago)

Quote:

cc2 said:
semaphore said:
Okay you didnt read it properly




of course, you couldn't be the one not explaining properly :rolleyes:

Quote:

semaphore said:
obviously i am not going to isolate or innoc from a contaminated dish :rolleyes: And its not contams of the mushroom culture itself its on the very outer edge some bacteria must have dropped onto the dish while i was working it




AFAIK contams. don't have to necessairly be over your mycelium for calling a plate contaminated. in fact most contams. unless it's cobweb will grow around your myc, not over it.

now let's also say bacteria are one of the easiest contams. to isolate healthy myc from, but it you had mold on the very outer edge it won't be so straightforward to inoculate with that.

if you've got some dishes that are contam-less, what's stopping you from inoculating then?





I was reading the same thing as well... That's why I didn't even reply


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Offlinesemaphore
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Re: Pan.Cyan jam agar plate, when to stop isolating. [Re: cc2]
    #14418117 - 05/08/11 05:38 AM (10 years, 8 days ago)

Quote:


if you've got some dishes that are contam-less, what's stopping you from inoculating then?




My question was not if i should inoculate, it was how many isolations should be done? i.e. what are the visuals i am looking for to know when i have a healthy culture growing for use. Sorry for my poor explanation its been a long two nights.


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Invisiblecc2
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Re: Pan.Cyan jam agar plate, when to stop isolating. [Re: cc2]
    #14418128 - 05/08/11 05:41 AM (10 years, 8 days ago)

Quote:

cc2 said:
if at 3rd isolation you're still getting contams. either isolate more or improve your sterile technique.




isolate until you haven't got any contams. on your plate.

you're looking only for mycelium nothing else.


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Offlinesemaphore
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Re: Pan.Cyan jam agar plate, when to stop isolating. [Re: cc2]
    #14418134 - 05/08/11 05:44 AM (10 years, 8 days ago)

Quote:

cc2 said:
Quote:

cc2 said:
if at 3rd isolation you're still getting contams. either isolate more or improve your sterile technique.




isolate until you haven't got any contams. on your plate.

you're looking only for mycelium nothing else.




Thanks, but that still doesnt answer my question. I am referring to the contam-less plates. With cubensis you get the sectoring, then you isolate that, let it grow then isolate again etc.. I was just wondering if it was a similar principle with cyans


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Invisiblecc2
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Re: Pan.Cyan jam agar plate, when to stop isolating. [Re: semaphore]
    #14418144 - 05/08/11 05:50 AM (10 years, 8 days ago)

depends on which features you're looking for.

if looking for fast colonization, isolate the fastest crawling mycelium, if for rope-like myc, isolate it etc.

however you can't predict carpophore characteristics from just mycelium. if you want a good isolate either cut out a ropey and fast one or fruit it and clone the best fruits.


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Offlinesemaphore
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Re: Pan.Cyan jam agar plate, when to stop isolating. [Re: cc2]
    #14418146 - 05/08/11 05:52 AM (10 years, 8 days ago)

Quote:

cc2 said:
depends on which features you're looking for.

if looking for fast colonization, isolate the fastest crawling mycelium, if for rope-like myc, isolate it etc.

however you can't predict carpophore characteristics from just mycelium. if you want a good isolate either cut out a ropey and fast one or fruit it and clone the best fruits.




cool thanks i have marked two plates that grew rather fast will try them out. Thanks for the info


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