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Offlinesoma_seeker
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100% non-colonization
    #5757802 - 06/16/06 03:51 PM (17 years, 7 months ago)

over the past 2 weeks i have inoculated at least a dozen jars. i initially inoculated to sets of four, of which 1 set were bright pink in 24hrs so i disposed of them, cleaned the jars, prep'd them again and inoculated.

again within 24hrs these jars were showing signs of pink contamination, (although FAR less than the previous jars) so once again i tipped them and tried again.

to date every jar i have inoculated except for 1 of the original set of 4 have showed at least a VERY small amount of pink contamination, but its not contamination i'm concerned about (otherwise i'd post in contam forum). i performed a control on my last set of jars by sterilizing (steaming) and then leaving them for 4 days to see if they contam and then didnt and i am now very stringent with my inoculation, flame sterilizing my needle after every inoculation site. this leads me to believe that the 3 diff sets of LC i've been using to inoculate must all be contam'd.

but as i said contamination isn't what is concerning me, what concerns me is that NONE of the jars (including the originals that show no signs of contam and were inoculated 10 days ago) have shown any signs of colonization whatsoever.

i'm confident i can beat the contams but i have no idea what is causing the jars not to colonise. i know it is a competition between the myc and the contams to colinate first but the amount of contamination in these jars is barely even visible and in some it seems to dissapear over time as well, but still no white spots visible.

any advice would be greatly appreciated.


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Taking a psychedelic is analogous to life, if you dwell on reaching the end you'll never enjoy the trip!


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InvisibleOptx
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Re: 100% non-colonization [Re: soma_seeker]
    #5757810 - 06/16/06 03:54 PM (17 years, 7 months ago)

get a new LC tek
or just use multispore


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please do not take everything i say here so personally. welcome to teh internets!


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Invisiblecappa
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Registered: 02/12/06
Posts: 854
Re: 100% non-colonization [Re: soma_seeker]
    #5758211 - 06/16/06 05:42 PM (17 years, 7 months ago)

Sounds like your LCs are hosed!


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Their are 10 types of people. Those that understand binary, and those who don't.
~Cappa.


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Offlinesoma_seeker
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Re: 100% non-colonization [Re: cappa]
    #5758241 - 06/16/06 05:52 PM (17 years, 7 months ago)

Quote:

cappa said:
Sounds like your LCs are hosed!




do u think that would explain the complete lack of growth though?
i mean even the small amount of contamination in my jars isn't growing...

note - i know to give up on a cake once it becomes contam'd, the only reason i'm leaving them is to c if any colonisation occurs. since i've retried the process several times i want to work out just how many problems are occuring in order to try and fix them.


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Taking a psychedelic is analogous to life, if you dwell on reaching the end you'll never enjoy the trip!


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Offlinekranked
it'smushtastic!!

Registered: 08/31/04
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Re: 100% non-colonization [Re: soma_seeker]
    #5758444 - 06/16/06 07:21 PM (17 years, 7 months ago)

I had a similair problem starting out. My jars didn't develope contam. but they also never colonized. My best guess is that I didn't allow the jars sufficient time to cool down before knocking them up. Although I don't even remember them being that hot either.


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"When it went off in New Mexico, that first atomic bomb, we thought of Alfred Nobel and his hope, his vain hope, that dynamite would put an end to all war"

-Robert Oppenheimer 1945


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OfflineCosmicFunGuy
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Re: 100% non-colonization [Re: kranked]
    #5758478 - 06/16/06 07:30 PM (17 years, 7 months ago)

I prep at least one extra jar so when its time to knock my jars up i can open the extra and put a thermometer inside to make sure that my cakes have sufficiently cooled. I then obviously throw this cake away. I save the jar for covering caps whilst making prints though. Definitely not needed but if you can't sit and be patient its a good way to go. I do it just as a precautionary measure.


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♫ ♪ In the garden of many fields, there is no me or you. There is no right or wrong. There is no up or down. There is no black or white. Everyone is the plow man. Everyone is the seed. Everyone harvests and everyone yields. In the garden of many fields, everyone is one with one love and one love is one with everyone. ♪ ♫
lyrics from: Transglobal Underground - Eyeway Souljah (from Psychic Karaoke)


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Offlinesoma_seeker
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Re: 100% non-colonization [Re: kranked]
    #5758482 - 06/16/06 07:30 PM (17 years, 7 months ago)

thanks kranked, but i'm leaving them at least overnight to cool and its quite cold where i am so i know thats not the prob.

i also just read (in the contam forum) that no mold will show up for at least 2 days and every instance of lipstick mold i've seen only occurs on already significantly colonised cakes.

i'm getting bright pink staining after only 12hrs for nearly every jar and no subsequent colonisation.

nothing i've tried has helped :frown:

i guess this probably does belong in the contam forum, i just didn't think contamination was the likely cause (newb)


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Taking a psychedelic is analogous to life, if you dwell on reaching the end you'll never enjoy the trip!


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Invisiblecappa
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Registered: 02/12/06
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Re: 100% non-colonization [Re: soma_seeker]
    #5758716 - 06/16/06 08:55 PM (17 years, 7 months ago)

If you have bacteria in your LCs, you won't see any molds grow in the jars for some time.

I'd put my money on bacteria infestation. Open a jar and smell it, that will let you know.


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Their are 10 types of people. Those that understand binary, and those who don't.
~Cappa.


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Offlinesoma_seeker
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Re: 100% non-colonization [Re: cappa]
    #5758812 - 06/16/06 09:25 PM (17 years, 7 months ago)

Quote:

cappa said:
I'd put my money on bacteria infestation. Open a jar and smell it, that will let you know.




thanks cappa. yeh the last one i threw out had only a very small amount of contam but smelt rancid.  :crazy:

so this explains the complete lack of myc growth?

guess i'll have to work harder on my LC prep sterilization rather than inoculation


--------------------
Taking a psychedelic is analogous to life, if you dwell on reaching the end you'll never enjoy the trip!


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Offlinesoma_seeker
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Re: 100% non-colonization [Re: soma_seeker]
    #5759554 - 06/17/06 01:16 AM (17 years, 7 months ago)

casualty number 10, 48hrs after inoculation (discarded)



:frown:  :confused:  :mad:


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Taking a psychedelic is analogous to life, if you dwell on reaching the end you'll never enjoy the trip!


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Offlinecoda
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Re: 100% non-colonization [Re: soma_seeker]
    #5760576 - 06/17/06 01:01 PM (17 years, 7 months ago)

remember that just because you see mycellium floating around in an LC doesn't mean it's cube mycellium. For all you know you could have cultured three lovely jars of lipstick molds. You already learned how to determine what the problem is (control jars), so i say whip up some new LC's and try again.


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To get really high is to forget yourself. And to forget yourself is to see everything else. And to see everything else is to become an understanding molecule in evolution, a conscious tool of the universe. And I think every human being should be a conscious tool of the universe. . . .

-JG

i really am glad you came back to us instead of taking the other path. *hug*

-A_S (RIP your final words to me will never be forgotten)



Don't fuck with the laughing jesus.


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Offlinesoma_seeker
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Re: 100% non-colonization [Re: coda]
    #5760957 - 06/17/06 03:59 PM (17 years, 7 months ago)

yeh thanks, guess i was kind of in denial of what the prob was. Think i will try multispore inoculation next.


--------------------
Taking a psychedelic is analogous to life, if you dwell on reaching the end you'll never enjoy the trip!


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Offlinecoda
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Re: 100% non-colonization [Re: soma_seeker]
    #5760998 - 06/17/06 04:12 PM (17 years, 7 months ago)

I've never really understood the thought behind starting an LC before growing out the culture. MS innoc is an MS innoc regardless of what it's innoculating. So, if you generate a shitty substrain you're going to have poor results. The only marginal benefit from starting from an LC is speedier colonization times.

Truthfully i think LC's should be done after one has determined what they want to keep and what they want to throw away. Who wants a qt jar of mycellium that produces weak potency, mutated mushrooms, and crappy flushes?

However hands on experience is great, and one should never be talked down to for trying new things or going a more advanced route there first time around. I would rather see newbies start out with grains and bulk and have difficulty then struggle with the PF tek. This whole hobby is about experimenting, and learning, and you really can't do either unless you go out on a limb sometime.

So even though your experiement failed you still learned some important lessons. Kudos man, better luck next time!


--------------------
To get really high is to forget yourself. And to forget yourself is to see everything else. And to see everything else is to become an understanding molecule in evolution, a conscious tool of the universe. And I think every human being should be a conscious tool of the universe. . . .

-JG

i really am glad you came back to us instead of taking the other path. *hug*

-A_S (RIP your final words to me will never be forgotten)



Don't fuck with the laughing jesus.


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Offlinesoma_seeker
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Re: 100% non-colonization [Re: coda]
    #5761670 - 06/17/06 07:55 PM (17 years, 7 months ago)

cheers coda. after my several failed attempts i'm just keen to have any method i try succeed. it's been quite disheartening (is that a word?) opening up the incubator each day after inoculation to be greated by more dam pink shit!

but yeh, patience and percervearance are virtues not to be taken lightly i guess. i'll get it yet!


--------------------
Taking a psychedelic is analogous to life, if you dwell on reaching the end you'll never enjoy the trip!


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