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Invisibled0urd3n
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Re: Rye Soak Water Liquid Culture! [Re: Pastywhyte]
    #21207641 - 01/31/15 04:53 PM (9 years, 11 months ago)

The fact that you guys get contams with LC when you have nocced up 1000's of jars probably scares me a bit.

But I am going to make my first LC this week anyways. :wink:


Don't really want to go through all 10 pages. If I rinse my wbs like 6-7 times like usual and soak for 24 hours that should work fine, right?


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OfflinehamloafM
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Re: Rye Soak Water Liquid Culture! [Re: d0urd3n]
    #21208907 - 01/31/15 09:12 PM (9 years, 11 months ago)

First flush from the first tub featuring the Rye GWLC in the original post.



Usually don't let fruits get so far along, but have been pretty busy as of late. 

The flush from the second tub ready to be harvested was as far along as the first one, so the decision to make prints was made.





Looks like another giveaway in the Marketplace is imminent.

Here goes a glimpse of the end of a 2 hour long research session with the WBS GWLC, circular magnetic stirbar, magnetic stir plate, gallon sized media vessels, and 2 inch magnetic stirbars. 



It was discovered that lining up the magnets in the gallon sized media vessels is going to be tricky.  The reason why is because there's a pretty deep canyon from the sides of the jar to the middle where the stirbar likes to rest.  The magnets have a hard time lining up under all that pressure of the liquid media, so a technique by the researcher HAD to be developed. 

Said technique involves finding the stirbar on the inside of the media, and tilting the jar side ways, holding the media vessel lid with gloved up, alcohol soaked hands, and centering the magnets.  Giving a kind of twirl action while simultaneously jerking the jar towards yourself, while at the same time, NOT get the media filtering material wet.  Developed said technique, achieved it twice after mapping out the motions in the mind, then decided to carry on with the sterilization of the media. 

The 2 inch stirbars were used in the gallon sized media vessels, the circular stirbars have found a home in quart jar sized media vessels, and once inch stirbars have found a home in pint sized liquid media vessels.  The mediums were then place in the 75x and sterilized for an hour and half at 15-17 psi, and are sitting safely in the pressure sterilizer below atmospheric pressure.


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InvisibleGrey
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Re: Rye Soak Water Liquid Culture! [Re: hamloaf]
    #21208960 - 01/31/15 09:25 PM (9 years, 11 months ago)

I'm about to build a stir plate. How often do you run it, and for how long?


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OfflinehamloafM
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Re: Rye Soak Water Liquid Culture! [Re: Grey]
    #21209683 - 02/01/15 04:12 AM (9 years, 11 months ago)

Not sure yet.  This is the first time researching with a magnetic stirbar and stirplate.

Seems to me that the device will only need to be ran for 30-60 seconds on the lowest rpm's possible whenever the cultivator feels it's necessary.  I plan letting the myce fill up the media, run the stirplate, let the culture settle, and once it fills the media again, stir it magnetically until a super highly concentrated liquid myclial culture is generated that has filled the media to the brim, aspire the culture, give it away in the Marketplace, then GET THE FUCK OUTTA' DODGE!

EDIT to add ~
I am only an avid LC researcher.  Figure why waste something you have 5 years experience in?  I only post my experiments, what I am researching, and my developments on the shroomery.  :wink:  :uptosomething:

The staples to my cultivation procedures are agar to grain.  G2G, then spawn to bulk.  I would like to replace the whole grainmastering thing with LC's one day though as a staple in the cultivation regimen. 


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Edited by liquidmyce (02/01/15 05:40 AM)

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InvisibleGrey
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Re: Rye Soak Water Liquid Culture! [Re: hamloaf]
    #21210381 - 02/01/15 10:17 AM (9 years, 11 months ago)

Haha, thanks. Good stuff, I'll be tuned in.


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OfflinehamloafM
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Re: Rye Soak Water Liquid Culture! [Re: d0urd3n]
    #21210471 - 02/01/15 10:46 AM (9 years, 11 months ago)

Quote:

d0urd3n said:
The fact that you guys get contams with LC when you have nocced up 1000's of jars probably scares me a bit.

But I am going to make my first LC this week anyways. :wink:




If THOUSANDS of jars have been lost to bacterial contamiantion, then have had MILLIONS of successful grows using LC's.  :super:

The only way to advance your/in liquid culturing techniques is to take the plunge, and begin field research study with them.  Rome wasn't built in a day.  Start small, and have several, successful small-scale grows using LC's before advancing your liquid culturing techniques, and procedures. 

Quote:

d0urd3n said:
But I am going to make my first LC this week anyways.




Luckily for you and others, LC isn't rocket science, and only takes a few times around the world (usually met with failure) to even begin to understand how they work for liquid culturing techniques really only have one specific use in this hobby therefore are an all-or-nothing artform of culturing.

Once you have your techniques, and procedures with LC's down pact, though, LC's are the ONLY way to expand your mycelial cultures by 10,000's as opposed the 1's of thousands that can be only be achieved using G2G transferring techniques.  As we ALL know (or may not know), though, G2G transferring techniques are the preferred method of procedures to begin with.  G2G transferring techniques are extremely noob-friendly, and basically bullet-proof, and is a staple procedure in this cultivators many tools of production techniques. 

Quote:

d0urd3n said:
Don't really want to go through all 10 pages. If I rinse my wbs like 6-7 times like usual and soak for 24 hours that should work fine, right?




Why, because you might learn something??  :confused:


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Invisibleblindingleaf
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Re: Rye Soak Water Liquid Culture! [Re: hamloaf]
    #21210479 - 02/01/15 10:49 AM (9 years, 11 months ago)

Ham, what's ur take on glc vs agar wedge> nutrient liquid?
Just curious what u like more and why


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OfflinehamloafM
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Re: Rye Soak Water Liquid Culture! [Re: blindingleaf]
    #21210536 - 02/01/15 11:10 AM (9 years, 11 months ago)

Comparing agar to LI, GLC, or liquid media, is like comparing apples and oranges.  There is no comparison.  Team agar ALL the way!.

GLC, and LI are considered the same thing.  Liquid inoculants aren't considered culture either though.  Again, no premeditated, nutrified media was applied, UNLESS you want to consider the amount of dissolved solids in any given water at any given time a media.  With those methods of inoculation, the only use of the water is a transferring medium for the mycelium.

The ONLY ways that are bullet proof to inoculate LC's with are by using a known clean agar culture, using either the "wedge and transfer" method, or to make a slurry concoction out of said known clean agar culture, aspire into syringes, or "free-pour" the concoction into your pre-sterilized, nutrified liquid media in front of a flowhood.


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OfflinehamloafM
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Re: Rye Soak Water Liquid Culture! [Re: Grey]
    #21210573 - 02/01/15 11:20 AM (9 years, 11 months ago)

What happens to the fruits when fruited without the polyfil is highly preferred over fruiting with the polyfil left in environmental-wise. 

Not trying to say that one method of fruiting deserves merit over the other, just that, in this cultivators experience, and opinion, the fruits generated when maximum airflow is allotted coupled by control of the humidity levels of the air in the environment are controlled produces a MUCH meatier, more golden colored stem, thicker capped, non-fuzzy-footed, heartier fruit.

However, the goal of this cultivator is to gain MAXIMUM use of the time, space, and materials available.  Gaining maximum use of the time, space, and materials available to you though takes a higher degree of, skill, will, work, determination, time, and force, AND (in my very strong opinion), said levels of degree of skill-sets are what separates the men from the boys, and the women from the girls .  If your goal is to utilize a set-and-forget method of mushroom production, then stuffing your monotub's holes with polyfil is for you.  More power to you.

This is one of the minimonos of the original Rye GWLC featured in the OP after having the polyfil completely removed for just 10 hours. 

 

Here goes a few more LC grows finished with their first flush while the polyfil was copletely stuffed into the holes for the duration of first flushing. 

Minimono featuring culture genrated by the original rye GWLC.
   

A couple of first flushed tubs inocualated with a LC inoculated with 3rd transfer from clone mycelium. 
 

Top tub.


Bottom tub.


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Invisibleeatyualive
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Re: Rye Soak Water Liquid Culture! [Re: hamloaf]
    #21211583 - 02/01/15 03:32 PM (9 years, 11 months ago)

those caps have a nice color! great work.
:rockon:


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OfflinehamloafM
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Re: Rye Soak Water Liquid Culture! [Re: eatyualive]
    #21218770 - 02/02/15 10:27 PM (9 years, 11 months ago)


Got out the WBS EOTBCGW, thawed it out, and loaded it into two gallon sized media vessels with 2 inch magnetic stir bar, and 1 quart jar with 3.5 circular magnetic stirbar, and Whatman syringe filter GE exchange filtration system modification.  Intended on diluting the media down to a 1:4 ratio (grain water:water), but somehow all of the jar's contents appear to be at different nutrient richness levels.

:shrug:

Sterilized gallon jar media 1.


Gallon jar media 1 is the richest with the most sediment in it, and therefore, as a result, is almost opaque.

Quart jar media 1.


Quart jar media 1 is the least richest, with the least amount of sediment in it, therefore, it's the most clearest than the other two mediums in it's appearance.

Gallon jar media 2.


Gallon jar 2 is a happy middle of richness in comparison and contrast to it's sister mediums.

All of thee above mediums were sterilized at 15 psi for an hour and a half.

It's becoming preferred to dilute the GW mediums using a 1:4 part grain water/water ratio just so growth can be observed.  Rye is pretty clear though without being diluted.  Plus, the GWLC generated by the undiluted rye when inoculated to sterilized rye berries was bad-ass all around (at mycelium running & at fruiting). 

Onto the undiluted high quality Red Milo liquid culture.

This medium is so DAMN rich that you can't see SHIT through this medium.  :mad2:



It's all good though, something is happening in there because mycelial growth has been noted to begin to form a mat over the top of the culture. 



That means something is alive in there, and am willing to bet it's a live, clean liquid mycelial culture.  The culture is scheduled to be aspired into a syringe or two to see what the hell is going on. 



IF a happy vibe is had, there await several sterilized red milo grain mediums to inoculate with this culture. 

The diluted red milo LC is still inside of a 60cc syringe eagerly wating to inoculate some red milo and WBS. 



Thanks for tuning in.  more to come!  :popcorn:


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Invisibled0urd3n
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Re: Rye Soak Water Liquid Culture! [Re: hamloaf]
    #21218781 - 02/02/15 10:29 PM (9 years, 11 months ago)

How do you get the LC out of those jars? Do you just tilt it or something?

Will be making some LC tomorrow I will post once I see some growth.


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OfflinehamloafM
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Re: Rye Soak Water Liquid Culture! [Re: d0urd3n]
    #21220289 - 02/03/15 10:06 AM (9 years, 11 months ago)

With one of these.  A stainless-steel liquid inoculation gun.  Had a good week at work this week, so just placed the order for the device.  It's well deserved.



Honestly, been asking the self the same thing.  Will most likely end up aspiring as much of the culture into syringes as I can without cross-contaminating, and give the culture away in the Marketplace considering the decision is made to inoculate these giant mediums with edible culture.

The major point of creating, and generating LC in a gallon sized media vessels was to research with the method of culturing in media vessels this size with a magnetic stirplate and magnetic stirbar rather than intending a grow with the culture, but I am not one for wasting, so with the culture left over after aspiring as many syringes as possible, the plan is to create sterilized substrates out of bulk substrate materials, inoculating them with generous amounts of LC, and fruiting inverto. 

The question is what to inoculate those giant liquid mediums with.  The options are (all options are known to be clean and have been cultured on agar) Cubensis, Blue Oyster culture, Pan stip, or atl#7.  What say ye?


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Invisibled0urd3n
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Re: Rye Soak Water Liquid Culture! [Re: Pastywhyte]
    #21221183 - 02/03/15 01:33 PM (9 years, 11 months ago)

Quote:

Pastywhyte said:
Quote:

hamloaf said:
Nice, Pasty.  What size jars are those, and how did you inoculate the media. 

You wrapped the SHIT out of those the lid seals with the parafilm, didn't you.  :lol:




Its a quart jar and it was inoculated with an agar wedge. I had a pin pop up on a plate and I transferred for a clean clone. Then when that grew out I inoculated this LC and a slant :thumbup:

And yes I did wrap the shit outta it with parafilm. What I lack in equipment I make up for in good choices and paranoia :wink: :paranoid:





It seems to me the main risk of LC (assuming you have good procedure and use agar) is the fact that when you stick your needle in the silicon port you can't sterilize the silicon, you can only wipe it down with iso. Which is why people say use large syringes (maybe one of you? I read too many LC threads lately). It seems to me like a contaminate getting through the lid would be the least of your worries. I guess better safe than sorry with LC, but it doesn't make much sense to me. Are contams getting through the lid more common that I had thought? Unless it is really wet I wouldn't think it would be an issue?

Thanks.


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Re: Rye Soak Water Liquid Culture! [Re: d0urd3n]
    #21221506 - 02/03/15 02:50 PM (9 years, 11 months ago)

My first and only GWLC - made in a baby food jar with no filter at all and done in a ghetto sab.
I have 10 jars of oats and bulk subs going ham from that LC now..
Of course I'm not advocating everyone use a ghetto sab and no filters or anything - but it works and I find people wayyyyyyu over analyze things and are overly safe/paranoid when it comes to cultivation

Its good to be safe and everything esp if u have all kinda of $ to spend on equipment, I'm just happy things aren't as complicated as the shroomery cult board makes them appear to be

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Re: Rye Soak Water Liquid Culture! [Re: d0urd3n]
    #21223510 - 02/03/15 09:33 PM (9 years, 11 months ago)

Quote:

d0urd3n said:
It seems to me the main risk of LC (assuming you have good procedure and use agar) is the fact that when you stick your needle in the silicon port you can't sterilize the silicon, you can only wipe it down with iso. Which is why people say use large syringes (maybe one of you? I read too many LC threads lately). It seems to me like a contaminate getting through the lid would be the least of your worries. I guess better safe than sorry with LC, but it doesn't make much sense to me. Are contams getting through the lid more common that I had thought? Unless it is really wet I wouldn't think it would be an issue?

Thanks.



Nope.  The main risk with LC's is inoculating them with spores, and nope.  The reason large syringes are best used for LC's is for the drawing of the culture into syringes from it's original media vessel. 

Large syringes are best used to aspire LC because live liquid mycelial culture is thick and will give you a hard time of you try to aspire it with an 18 guauge needle.

Pushing contams that may be on the injection port into the media with the needle of the syringe upon inoculation is also a risk factor when inoculating LC's with a syringe through an injection port.


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Invisibled0urd3n
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Re: Rye Soak Water Liquid Culture! [Re: hamloaf]
    #21223977 - 02/03/15 11:45 PM (9 years, 11 months ago)

Do you just not use an injection port then? I see not all of your lids have them. You think that is safer than using an injection port to just unscrew the lid? I hope 18 ga works alright, it is all I have right now.


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Re: Rye Soak Water Liquid Culture! [Re: d0urd3n]
    #21228395 - 02/04/15 10:09 PM (9 years, 11 months ago)

The saga continues!

Here goes the undiluted Red Milo LC just before being shook.



After rigorously shaking the culture, growth, one centimeter thick was FINALLY observed just below the surface of the media.



What culture was available was aspired into syringes



and used to inoculate sterilized Red Milo mediums. 



Inoculated those gallon, and quart jar WBS GWLC mediums with 1 cc of Blue Oyster Mushroom mycelium a piece.



The PESH LC's made last week with WBS GWLC, and magnetic stirring capacities were stirred up using the magnietic stirplate this evening. 

First, the WBS GWLC in the quart sized media vessel containing 3.5 circular magnetic stirbar was stirred.  Started the magnetic stirplate at 200 rpms, and slowly increased the rpm speed until the magnetic stirbar generated a cyclone in the media strong enough to start shredding/processing the culture.  The stirbar it's self played a major role in the processing/shreeding of the culture, as well.

The anomaly of the stirbar and plate creating enough cyclonic force to begin shredding the culture took place at about 600 rpm's.  The rpm speed was increased a little, and topped out at 675, and was sustained for about 120 seconds, then the device was turned off.

   

The WBS GWLC in the pint sized jar containing the 1 inch magnetic stirbar was mixed next.  The media in the pint jar with 1 inch magnetic stirbar took a much greater amount of rpm's before the culture began to become processed by the stirbar it's self, and cyclonic force generated by said stirbar.

   

Looking forward to more research with this magnetic stirplate and magnetic stirbar.  The precision at which these devices breaks up, and stirs the liquid mycelium is great.  The added bonus of not having to shake the culture by hand is a pleasant delight, and a dream come true also.

Can't wait to get the hands on that brand new, stainless-steel, 10cc capacity inoculation gun, and begin practicing bulk LC inoculations on a small scale until a few times around the world is gone using bulk liquid culturing techniques that are met with success before consideration is even made to replace grain to grain transferring techniques with liquid culturing techniques as a staple procedure in the exponential expansion of mushroom mycelium.  Thanks for tuning in.  So much more to come.

GWLC MANIA!


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Re: Rye Soak Water Liquid Culture! [Re: hamloaf]
    #21228402 - 02/04/15 10:11 PM (9 years, 11 months ago)

Hey. Where'd you end up getting that stirrer?  You like?


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Re: Rye Soak Water Liquid Culture! [Re: azur]
    #21228484 - 02/04/15 10:31 PM (9 years, 11 months ago)

Quote:

azur said:
Hey. Where'd you end up getting that stirrer?  You like?




Yeah, curious as well. How are you gonna clean the inoculation gun? I was looking at that and it said it couldn't be autoclaved.


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