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OfflineDiosocragenes
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Registered: 01/28/18
Posts: 52
Last seen: 6 years, 8 months
Need LC Advice
    #25154040 - 04/20/18 07:00 PM (6 years, 8 months ago)

I started my first LC a few days ago, there's a cloud building up. I followed the LC cloning pictorial tek and took tissue from a PEU fruit. Now in the tek it didn't mention anything about putting a marble/shard of glass in the LC jar to break up the mycelium when it starts getting thick. I had stumbled across using a marble/glass after completing the tek. For the people that have experience with LC, do you recommend sterilizing a marble/glass and then inserting into my LC jar in a glove box? Or is that too risky for contamination and stirring it a few times a day it will be ok? Thanks for any help always appreciated.

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OfflineChwyn
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Registered: 08/21/16
Posts: 1,238
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Re: Need LC Advice [Re: Diosocragenes]
    #25154046 - 04/20/18 07:07 PM (6 years, 8 months ago)

you could try that or whenever you go to take some LC just draw some of the liquid in the syringe and force it back out before drawing some in. The pressure will mix and break it up a bit.


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OfflinemyceliupsS
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Registered: 01/24/15
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Re: Need LC Advice [Re: Diosocragenes]
    #25154098 - 04/20/18 07:43 PM (6 years, 8 months ago)

Quote:

Diosocragenes said:
I started my first LC a few days ago, there's a cloud building up. I followed the LC cloning pictorial tek and took tissue from a PEU fruit. Now in the tek it didn't mention anything about putting a marble/shard of glass in the LC jar to break up the mycelium when it starts getting thick. I had stumbled across using a marble/glass after completing the tek. For the people that have experience with LC, do you recommend sterilizing a marble/glass and then inserting into my LC jar in a glove box? Or is that too risky for contamination and stirring it a few times a day it will be ok? Thanks for any help always appreciated.





Do you have a lot of experience in agar? If not I would start
there. Most people take their mushroom tissue and put to agar
first to clean it up. And don't put any marbles in there unless
there were sterilized with th LC.

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OfflineDiosocragenes
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Registered: 01/28/18
Posts: 52
Last seen: 6 years, 8 months
Re: Need LC Advice [Re: myceliups]
    #25154145 - 04/20/18 08:08 PM (6 years, 8 months ago)

I don't have any experience with agar. Still learning about it, plan to start that adventure soon.

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OfflineDiosocragenes
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Registered: 01/28/18
Posts: 52
Last seen: 6 years, 8 months
Re: Need LC Advice [Re: Chwyn]
    #25154154 - 04/20/18 08:12 PM (6 years, 8 months ago)

Quote:

Chwyn said:
you could try that or whenever you go to take some LC just draw some of the liquid in the syringe and force it back out before drawing some in. The pressure will mix and break it up a bit.




I think I'll try putting a sterile shard of glass in it, as soon as it's completely colonized ill test it out to see if it's good or not. Then do it the right way next time.

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OfflinemyceliupsS
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Re: Need LC Advice [Re: Diosocragenes]
    #25154155 - 04/20/18 08:12 PM (6 years, 8 months ago)

LC is something to do after you have mastered agar.
In most cases. I'm not saying it won't work, but a
cloud building up is typically a bad sign.

Read in detail EVERYTHING HERE!!!!

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OfflineDiosocragenes
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Registered: 01/28/18
Posts: 52
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Re: Need LC Advice [Re: myceliups]
    #25154160 - 04/20/18 08:16 PM (6 years, 8 months ago)

I thought as it started to colonize it would get a cloudy appearance... thanks for the link

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OfflineThedillestpickle
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Re: Need LC Advice [Re: Diosocragenes]
    #25154217 - 04/20/18 09:03 PM (6 years, 8 months ago)

there should be a cloudy clump that forms.  It will be formed of strands of mycelium.  It should fill about 1/3 of the volume of the liquid. 

I wouldn't put any marbles or shards in it.  I wouldn't open it to add anything either, as that would just make it more likely to contaminate. 

The best technique for making LC that I have come across is called "agar broth", it is somewhere buried in the older threads.  You might be able to find it through a search.

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OfflinehamloafM
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Re: Need LC Advice [Re: Thedillestpickle]
    #25154229 - 04/20/18 09:13 PM (6 years, 8 months ago)

The culture it's self will appear as a cloud, but you want the media to be see through.  If media is cloudy, or non transparent in any way LC is contaminated.

https://www.shroomery.org/forums/showflat.php/Number/21068298#21068298


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

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Offlinerosco420i
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Registered: 07/25/09
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Re: Need LC Advice [Re: hamloaf]
    #25154258 - 04/20/18 09:39 PM (6 years, 8 months ago)

I just started doing LC's and cloning from mushroom tissue recently. I've never used agar, this is the 1st time that I've done any type of cloning or liquid cultures and I have knocked up 14 jars with a 100% success rate. What I did was I found a magnetic stirrer on Amazon for $40 got magnetic stir pills, sterilize my LC with the stir pill in there and then put it on the magnetic stirrer and it creates a little tornado in there to constantly mix it as it's colonizing. If you don't have that or don't want to deal with it, you want to go in and shake them vigorously each day from what I've heard. But a magnetic stirrer is ideal. Also to break up the cloud you can order like 4" needles that can easily reach down into a quart jar and just move it back and forth and there to break it up a bit to suck some up in your syringe. Over all pretty easy to do, however  The long needles I bought, the part that screws into the syringe melted the 1st time I sterilized them so depending on which ones you find I would sterilize it with boiling water and not a pressure cooker


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"He who makes a beast of himself gets rid of the pain of being a man"
                                                                            - Hunter S. Thompson.

Edited by rosco420i (04/20/18 09:43 PM)

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