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Offlinemickle207
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Registered: 09/03/10
Posts: 164
Last seen: 4 years, 4 months
LC contamination?
    #14303877 - 04/16/11 11:08 PM (12 years, 10 months ago)

Ok so I made some LC using the easy clone tek where you just puncture the base of the stem with a needle and then inject it into a jar.
I inoculated many batches of bags and jars with it and all of them seemed to colonise well until about 70% and I start to have green mold which stunts the colonization.
I've tried re-doing batches over and over again and the results are the same. I have now a 100% failure rate and I doubt it was because of my sterile techniques since this never happened to me before. I'm just trying to narrow down the problem and I think the LC is the problem, but I'm not sure if one is using contaminated LC, will there be any growth at all?

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InvisibleDoc_T
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Re: LC contamination? [Re: mickle207]
    #14303897 - 04/16/11 11:12 PM (12 years, 10 months ago)

LC :thumbdown:


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Invisibleslapphappypill
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Re: LC contamination? [Re: Doc_T]
    #14304060 - 04/16/11 11:55 PM (12 years, 10 months ago)

LC can have growth with contams as well, it just doesn't seem to be strong enough to fight the contams. Next time just test a couple small jars first


--------------------
We think we have freedom, but we're all just mice in a maze....

                   

FYI: I stole all my pix off google! F+ PORN! Here is a shit-ton of porn by yours truly!

I have FINALLY written up a couple teks as to how SHP has done things in the past. DISCLAIMER: This is not for the newbie to mycology, and not going to work for everyone! This is simply what works for one person when other teks and methods have failed miserably!
~~~~~How SHP does their unconventional WBS Prep!! (NO DRY METHOD)~~~~
~~~~~SHP's highly disputed method of doing ALL their work outside of a flow hood or a Still air box!~~~~~
~~~~~Troubles harvesting the side and bottom pins in your mono? Learn how!! Dunking included ;-)~~~~~

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Offlinemickle207
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Registered: 09/03/10
Posts: 164
Last seen: 4 years, 4 months
Re: LC contamination? [Re: Doc_T]
    #14304062 - 04/16/11 11:55 PM (12 years, 10 months ago)

I'll take that that my LC is contaminated...
So I should move on and use agar right?

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InvisibleJavadog
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Re: LC contamination? [Re: mickle207]
    #14304266 - 04/17/11 01:05 AM (12 years, 10 months ago)

If you can, definitely.


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Boyd Rice told my brother that life is a corny pack of freesakes

Myco-tek.org

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Invisibleslapphappypill
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Re: LC contamination? [Re: Javadog]
    #14304279 - 04/17/11 01:08 AM (12 years, 10 months ago)

Yeah, Agar is the way to go for sure! However for me, I still use agar to LC and works perfect for me. If you can get it down right with the proper sterile procedure, LC is perfect for grain jars and bags IMO

The reason LC gets such a bad rep is because people like to jump into tones of jars from a LC that wasn't tested. If you have a clean LC, it can be your absolute best friend especially if you don't have a flow hood!


--------------------
We think we have freedom, but we're all just mice in a maze....

                   

FYI: I stole all my pix off google! F+ PORN! Here is a shit-ton of porn by yours truly!

I have FINALLY written up a couple teks as to how SHP has done things in the past. DISCLAIMER: This is not for the newbie to mycology, and not going to work for everyone! This is simply what works for one person when other teks and methods have failed miserably!
~~~~~How SHP does their unconventional WBS Prep!! (NO DRY METHOD)~~~~
~~~~~SHP's highly disputed method of doing ALL their work outside of a flow hood or a Still air box!~~~~~
~~~~~Troubles harvesting the side and bottom pins in your mono? Learn how!! Dunking included ;-)~~~~~

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Offlinemickle207
Stranger Shrooms

Registered: 09/03/10
Posts: 164
Last seen: 4 years, 4 months
Re: LC contamination? [Re: slapphappypill]
    #14304572 - 04/17/11 03:13 AM (12 years, 10 months ago)

So... I have no idea how to work with agar yet, but from what i've read, it seems that u can just cut out contaminated parts?
If so, can I use my lc to inoculate to agar and cut out the contaminated parts? If I can that would be great since this LC is the ONLY remaining sample I have now... If I can't then i'll have to wait 2 weeks until my order ships from sporeworks.....

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Invisibleslapphappypill
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Re: LC contamination? [Re: mickle207]
    #14304613 - 04/17/11 03:39 AM (12 years, 10 months ago)

yes, you can knock up an agar dish with LC and pull the healthy myc out


--------------------
We think we have freedom, but we're all just mice in a maze....

                   

FYI: I stole all my pix off google! F+ PORN! Here is a shit-ton of porn by yours truly!

I have FINALLY written up a couple teks as to how SHP has done things in the past. DISCLAIMER: This is not for the newbie to mycology, and not going to work for everyone! This is simply what works for one person when other teks and methods have failed miserably!
~~~~~How SHP does their unconventional WBS Prep!! (NO DRY METHOD)~~~~
~~~~~SHP's highly disputed method of doing ALL their work outside of a flow hood or a Still air box!~~~~~
~~~~~Troubles harvesting the side and bottom pins in your mono? Learn how!! Dunking included ;-)~~~~~

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Invisiblehamloaf
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Registered: 12/23/09
Posts: 20,584
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Re: LC contamination? [Re: mickle207]
    #14304686 - 04/17/11 04:44 AM (12 years, 10 months ago)

Quote:

mickle207 said:
So... I have no idea how to work with agar yet, but from what i've read, it seems that u can just cut out contaminated parts?
If so, can I use my lc to inoculate to agar and cut out the contaminated parts? If I can that would be great since this LC is the ONLY remaining sample I have now... If I can't then i'll have to wait 2 weeks until my order ships from sporeworks.....



Like slappy said, LC can be used to inoculate sterilized agar with. 


When cleaning up cultures on agar, don't cut the contaminated parts of the culture out.  Cut out clean myce tissue samples from the very leading edge of the culture, transferring the healthy mycelium away from the contamination onto freshly sterilized agar in petri dishes.  Used to do this inside of a still-air box.  Now healthy myce is transferred away from contaminates inside of a clear plastic trash bag.  Next time cultures need to be cleaned from contamination, plan on using the hot agar pour as the means to clean the culture.  That way cultures can be cleaned in front of the flowhood. 


Gain a fast idea of how to work with agar by going to RR's profile and clicking on the mushroom sample videos link in his sig...  There you'll find a short instructional video over how to prepare agar (along with several other instructional sample videos) I think you may find helpful.


--------------------

   
How I Get Stuff done. - My Reference Guide. - My Grows.

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Offlinemickle207
Stranger Shrooms

Registered: 09/03/10
Posts: 164
Last seen: 4 years, 4 months
Re: LC contamination? [Re: hamloaf]
    #14310424 - 04/18/11 03:58 AM (12 years, 10 months ago)

Wait... i just thought of something and have been browsing abt it in the contamination forum. All the jars i've used were previously contaminated and i only used sunlight to clean it out thoroughly. is that sufficient for cleaning jars? I thought it would be since it will go into the pressure cooker anyways. But, the same type of green mold also happened to my bags and I have never reused a bag before.

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