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Offliner00tuuu123
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Re: Rye Soak Water Liquid Culture! [Re: shroominmyroom]
    #21151267 - 01/20/15 05:41 PM (9 years, 11 months ago)

Quote:

shroominmyroom said:
Quote:

r00tuuu123 said:
That's just after the boil just wbs tea so to speak. The jars are in the pc right now about 15 minutes left then have to let them cool. Will probably noc them in the am. :wink: Hopefully this will work as I am doing it old school with a marble in the jar and not an electric stirrer.




I don't like marbles in LC. Doesn't do jack shit.



Well that's what I got to work with so I guess it'll have to do. :shrug:


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Re: Rye Soak Water Liquid Culture! [Re: r00tuuu123]
    #21151356 - 01/20/15 06:04 PM (9 years, 11 months ago)

I used to do the broken glass or screws  but now I don't bother.  A good swirl usually does the trick for me.

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Re: Rye Soak Water Liquid Culture! [Re: Pastywhyte]
    #21151362 - 01/20/15 06:05 PM (9 years, 11 months ago)

I just got better syrines lol


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Re: Rye Soak Water Liquid Culture! [Re: cronicr]
    #21151386 - 01/20/15 06:10 PM (9 years, 11 months ago)

Today I used that LC  I posted. No marble glass or anything and I had no problems after shaking hard - should have taken pics of the syringe but it was beautiful, like 5cc half the syringe  of straight myc :hellyeah:

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Offliner00tuuu123
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Re: Rye Soak Water Liquid Culture! [Re: Goldberg]
    #21151417 - 01/20/15 06:16 PM (9 years, 11 months ago)

Quote:

Goldberg said:
Today I used that LC  I posted. No marble glass or anything and I had no problems after shaking hard - should have taken pics of the syringe but it was beautiful, like 5cc half the syringe  of straight myc :hellyeah:



Yea but I'm working from spores not slurry. :hmm:


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OfflineGoldberg
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Re: Rye Soak Water Liquid Culture! [Re: r00tuuu123]
    #21151476 - 01/20/15 06:32 PM (9 years, 11 months ago)

Sorry I wasn't responding to anythig you said just telling everybody I didn't need anythig in my jar to break the myc up, but not saying that doesn't help

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OfflinehamloafM
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Re: Rye Soak Water Liquid Culture! [Re: r00tuuu123]
    #21153895 - 01/21/15 05:50 AM (9 years, 11 months ago)

Quote:

r00tuuu123 said:

Yea but I'm working from spores not slurry. :hmm:



Oooooh, inoculating liquid media with spores are no-goods.

Hope everything goes well for you, but you are taking a HUGE gamble that your media will contaminate inoculating the media with spore syringe solution.  Good luck!

Even IF your media contaminates I would ask, AND encourage you to post your results here.  Let us know how it goes, please.  Thanks.

I use 2 broken glass shards in my LC's to break them up, as well.  I VIGOROUSLY shake the shit out of them though just prior to aspiration.  Usually after the culture has been shook, and I'm cleaning out the media vessel, I'll find that the 2 shards of glass have multiplied.  Can't wait to begin developing research with the magnetic stirplate and stirbar. 

GOLDBERG, Your LC looks good, man.  You inoculated the media the right way too.  Good job!

Let us know how your liquid culture develops!


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Re: Rye Soak Water Liquid Culture! [Re: hamloaf]
    #21153976 - 01/21/15 06:49 AM (9 years, 11 months ago)

I just PC'd me up some wheat gw and plan to inoculate with a good looking clone I want to test. Dilution was 1:2 but it still looks quite rich, pics to follow :thumbup:

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OfflinehamloafM
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Re: Rye Soak Water Liquid Culture! [Re: Pastywhyte]
    #21153979 - 01/21/15 06:51 AM (9 years, 11 months ago)

Nice!  Thank you for your cooperation, and continued support, Pasty.  Appreciate it.  :thumbup:


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Offliner00tuuu123
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Re: Rye Soak Water Liquid Culture! [Re: hamloaf]
    #21153997 - 01/21/15 07:00 AM (9 years, 11 months ago)

Quote:

hamloaf said:
Quote:

r00tuuu123 said:

Yea but I'm working from spores not slurry. :hmm:



Oooooh, inoculating liquid media with spores are no-goods.

Hope everything goes well for you, but you are taking a HUGE gamble that your media will contaminate inoculating the media with spore syringe solution.  Good luck!

Even IF your media contaminates I would ask, AND encourage you to post your results here.  Let us know how it goes, please.  Thanks.

I use 2 broken glass shards in my LC's to break them up, as well.  I VIGOROUSLY shake the shit out of them though just prior to aspiration.  Usually after the culture has been shook, and I'm cleaning out the media vessel, I'll find that the 2 shards of glass have multiplied.  Can't wait to begin developing research with the magnetic stirplate and stirbar. 

GOLDBERG, Your LC looks good, man.  You inoculated the media the right way too.  Good job!

Let us know how your liquid culture develops!



Like I said in a previous post I'm working with what I have on hand currently I'm not set up to do agar and money is gonna be tight till april. I've done lcs with spores in karo or honey mixes but they did not perform to the levels I expected they seemed to stall out after a while. I'm hoping that the WBS water has that certain something I am missing.


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Re: Rye Soak Water Liquid Culture! [Re: r00tuuu123]
    #21154009 - 01/21/15 07:05 AM (9 years, 11 months ago)

Compared to using sugar LC's, I am SURE that your WBS GWLC will WAY out perform those aforementioned sugar LC's.

Upgrading your sterile procedures and techniques will give you that OTHER something you are missing.

There are plenty of cheap, non-flowhood having ways of doing agar.  Pasty's Easy Agar Tek is a GREAT example. 

Gas Exchange issues are the most common reason for LC's to stall.

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Re: Rye Soak Water Liquid Culture! [Re: hamloaf]
    #21154116 - 01/21/15 07:56 AM (9 years, 11 months ago)

Quote:

hamloaf said:
Compared to using sugar LC's, I am SURE that your WBS GWLC will WAY out perform those aforementioned sugar LC's.

Upgrading your sterile procedures and techniques will give you that OTHER something you are missing.

There are plenty of cheap, non-flowhood having ways of doing agar.  Pasty's Easy Agar Tek is a GREAT example. 

Gas Exchange issues are the most common reason for LC's to stall.



I used to use a home made pressurised glove box quite simple in its construction rubber gloves white trashbags for sleves with duct tape a piece of plexiglass siliconed to the bottom which was then the top so I could see what I was doing a hepa filter on one end fed by a small space heater  and a bunch of 1/4 inch holes on the other. My sterile teknique is not an issue but contams were never a problem. I'll look into the gas exchange problem Right now using postal tyvek for that What do you think Maybe switch to pollyfill or filter disks on my next run?:shrug: Oh and BTW the reason I'm not doing agar right now is I just don't have the coin to lay out for it right now.


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Re: Rye Soak Water Liquid Culture! [Re: r00tuuu123]
    #21154181 - 01/21/15 08:19 AM (9 years, 11 months ago)

I prefer a whatman filter for LC, tyvek and polyfil will wick contams easily if they get wet. If you already have a SAB or clean workspace for sterile work then my agar tek costs less than $20 to do. Not sure if that is outside your budget but its one of the cheapest ways to start with agar.

Also regardless of success rates a simple SAB is very effective to work in and will cost a lot less than a positive pressure setup.

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Re: Rye Soak Water Liquid Culture! [Re: Pastywhyte]
    #21154227 - 01/21/15 08:36 AM (9 years, 11 months ago)

Quote:

Pastywhyte said:
I prefer a whatman filter for LC, tyvek and polyfil will wick contams easily if they get wet. If you already have a SAB or clean workspace for sterile work then my agar tek costs less than $20 to do. Not sure if that is outside your budget but its one of the cheapest ways to start with agar.

Also regardless of success rates a simple SAB is very effective to work in and will cost a lot less than a positive pressure setup.



Thanks man that is encouraging cost wise right now i'm living hand to mouth time to dig in the couches and get enough to buy petris and agar and hit up one of the print lotteries in the trade forum. please link your tek.


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Re: Rye Soak Water Liquid Culture! [Re: r00tuuu123]
    #21154276 - 01/21/15 08:43 AM (9 years, 11 months ago)

Pasty's agar tek is here
http://www.shroomery.org/forums/showflat.php/Number/19208976

Since expense is an issue for you, you could consider using screw-top containers.  They are a bit more expensive each but have a very wide variety of uses so I think they go further per dollar.
http://www.shroomery.org/forums/showflat.php/Number/19035259


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Re: Rye Soak Water Liquid Culture! [Re: Pastywhyte]
    #21154285 - 01/21/15 08:45 AM (9 years, 11 months ago)

Quote:

Pastywhyte said:
I prefer a whatman filter for LC, tyvek and polyfil will wick contams easily if they get wet. If you already have a SAB or clean workspace for sterile work then my agar tek costs less than $20 to do. Not sure if that is outside your budget but its one of the cheapest ways to start with agar.

Also regardless of success rates a simple SAB is very effective to work in and will cost a lot less than a positive pressure setup.





Yeah, I use these lids for LC.



Oh, Pre made lids are $4.


These injection ports are 20 cents.



Oh, The LC filter is about $1.40.

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Invisibleshroominmyroom
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Re: Rye Soak Water Liquid Culture! [Re: blackdust]
    #21158116 - 01/21/15 03:51 PM (9 years, 11 months ago)

:flyhigh:

Edited by shroominmyroom (12/05/17 12:38 PM)

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Re: Rye Soak Water Liquid Culture! [Re: Pastywhyte]
    #21158148 - 01/21/15 03:59 PM (9 years, 11 months ago)

Quote:

Pastywhyte said:
I prefer a whatman filter for LC, tyvek and polyfil will wick contams easily if they get wet. If you already have a SAB or clean workspace for sterile work then my agar tek costs less than $20 to do. Not sure if that is outside your budget but its one of the cheapest ways to start with agar.

Also regardless of success rates a simple SAB is very effective to work in and will cost a lot less than a positive pressure setup.




After my limited work on petris I still plan to test out your easy agar tek to compare!

Question: do you use petris still or use the mini rounds for all your agar work?

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Re: Rye Soak Water Liquid Culture! [Re: Guardian187]
    #21158245 - 01/21/15 04:07 PM (9 years, 11 months ago)

I still use a little pour agar but not much. I just find pouring to be a major pain in my SAB. It can be done of course, its just not my preference. When/if I get a hood I will probably do mostly pour agar.

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Re: Rye Soak Water Liquid Culture! [Re: Pastywhyte]
    #21163870 - 01/22/15 04:53 PM (9 years, 11 months ago)

Quote:

I'll look into the gas exchange problem Right now using postal tyvek for that What do you think Maybe switch to pollyfill or filter disks on my next run?



For use as gas exchange filtering material, postal tyvek is about as worse as it gets.  I mean, it's good if you are in a pinch, but expect it to only work 1-2 times MAX.  Postal tyvek needs to be replaced every other run the pressurizer due to the intense heat of the pressurizer-cycle causing the pores of the materials to close up, HALTING gas exchange all together. 

As the guys suggested, Whatman filters are the best for LC's. 

When doing 1L or more worth of LC in a single media vessel, longer sterilization-cycles, and increased measures to gas exchange MUST be employed.

One of the peers of mushroom cultivation donated 3 Whatman filters to research the other day.  Lids have already been made out of the filters.



1 gallon of End Of The Boil-Cycle Grain Water (WBS) has been collected for liquid culturing media developmental research, and diluted to 50%, allowed to cool, then placed in the freezer for storage. 



:hehehe:


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