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InvisibleMudaFuka
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Liquid Inoculant ( NEW version) * 30
    #20429745 - 08/16/14 10:53 PM (6 years, 2 months ago)

Un like LC which contains nutrients and needs time to colonise. LI is made by mixing a colonised agar plate into sterile water. This is a real improvement because it greatly lowers the potential for contamination.

It also removes the need to do G2G transfers since you can simply make enough LI to inoculate as much spawn as you want right at the initial inoculation.

There are PC able blender jars on the market designed for this purpose but they are a bit pricey. So I made my own

Materials list
* An Oster blender ( They can be found very inexpensively and I'm not aware of a model that won't work for this tek)

* A regular mouth 1/2 pint mason jar with a metal lid

* Any 100% silicone (I prefer the clear kitchen and bath but I'll use whatever I have handy)


Making your blender jar

SIDE NOTE: The pictures in this portion of the tek were taken of two different lid assemblies. don't be confused by the different colored silicone and metal lids used.

Start by cutting the center out of your metal lid. I do this by drilling a hole in the center. Then using wire cutters or needle nose pliers to make the hole bigger. If you have some really good smaller tin snips they should work to but the lids are quite thin and cheep snips will just make a mess.



Now attach what's left of your jar lid to the blade side of your Oster blade assembly using 100% silicon. Be sure to sand both the lid and assembly before applying silicone or it will not properly adhere to either surface.



Now place the lid and assembly blade side down on your regular mouth 1/2 pint jar. I like to coat the jar's rim with dish soap to keep silicone from sticking to it.



It's time to make this a one piece lid. Run a bead of silicone under the lip of your jar's lid ring and screw it on to the jar. Be sure to sand both surfaces that will come into contact with the silicone.



Smooth down the excess silicon and leave the lid to dry for 24 hours.




After 24 hours the silicone on the top of the lid will have cured but the silicon on the inside of the jar will need more time in open air. Take your new blender lid off and let it sit until the silicone on the blade side has cured.



LI extraction lids

When it comes time to use your LI, you will want to extract it from the blender jar into a syringe. For this I use a modified plastic lid.


I make it by drilling 2 holes in a regular mouth plastic lid.

One hole at 1/4" diameter is filled with silicone. It is to be used as a self healing injection port or SHIP. I prefer clear silicone for the SHIP as it lets you see the hole underneath.


The other hole I make at 1/16" diameter. it gets covered with a small piece of synthetic filter disc or SFD. I affix the SFD using silicone. This second hole will allow filtered air to enter the jar. without it a vacuum is created and it is nearly impossible to draw out any liquid. 

Before applying silicone to a plastic lid. The lid must be sanded to remove the smooth finish.



If you don't have access to SFDs you can use tyvek 

This lid will need to be sterilised. Wrap it in foil and PC it along with your blender jar.



Making LI

This is the process of blending a colonised agar plate in sterile water to make inoculant.

Start out by filling your jar with water. I use 180ml in each jar for 1 colonised plat.


Put the lid on slightly loose, cover with foil and sterilise in your pressure cooker for 20 minutes at 15 PSI.



Once the PC cycle has ended and the pressure has dropped back down to zero on its own. Take out your jar tighten the lid and put it in your still air box or in front of your flow hood to cool. I usually put the plate I plan on using into my still air box at this point as well.



After a couple of hours the jar should be cool to the touch and ready to use. At this point prep your SAB by misting it heavily with soapy water to trap any contaminant on the SAB's floor and walls. Turn off any fans, close all your windows and give the air in your SAB time to settle. 


Now that the air has settled put on a pair of gloves and a face mask and get to work. Soak your gloves in 70% iso alcohol or a alcohol based hand sanitizer. Start by wiping down your jar and plate with a Lysol wipe or alcohol soaked paper towel. Loosen the jars lid but leave it on the jar.

You will need a scalpel or exacto blade to cut your colonised agar from the plate. Flame sterilise your blade. I find a propane or butane torch works well.



Cut out a circle of colonised agar. Make sure it is small enough to fit into your 1/2 pint jar without hitting the rim. While holding the colonised agar on the end of your scalpel. Take the lid off of your blender jar with the other hand and drop the agar into the jar. Watch that your hand are never directly above the open plate of jar. Screw the lid back on securely and it is ready to blend



To blend your agar. Simply hold your jar securely on to the blender base.


Do not blend to long or you risk damaging the mycelium and prolonging colonisation times. I have found 4 seconds at low to medium speed works well. If it is still chunky after the first 4 seconds I will give it 1 more second at high speed.



You should now have a jar of perfectly blended Liquid inoculant.



At this point you can use it to inoculate things like grain jars or bottle subs by simply pouring a small amount into each jar. For the sake of accurate even distribution of inoculant. I have started using 60cc syringes. I have found 16 gauge needles work best for this. 18 gauge are to narrow and 14 gauge damage the SHIPs



This is very simple. just put you blender jar back in to your SAB and wipe it off with a Lysol wipe or alcohol soaked paper towel. unscrew the lid but do not remove it yet. get your LI extraction lid ready. open the foil. Pick up the blender lid with one hand and quickly replace it with the plastic lid.


Sick a sterile needle into the SHIP and pull back on the plunger until the syringe is full.


You can fill 3 60cc syringes with each jar. I generally use them right away but they can be stored in the fridge for future use. I don't know exactly how long they will stay viable but I have used LI after more than a year in storage and they worked just fine.

Inoculation

If you did ever thing right you now have a few syringes full of LI ready to go



Start by loading all your jars, bags or whatever you plan on inoculating into your Still air box. At this point I also put my LI syringes into the SAB.



Prep just like before by misting your SAB with soapy water, turning off fans, putting on gloves and waiting for the air to settle.



Once the air has settled. Wipe down all your jar lids and your syringes with a Lysol wipe or alcohol soaked paper towel.



Flame your needle.



Many of my Jar lids have the same type of silicon self healing injection ports as my Li extraction lid. So I just stick my needle into the SHIP and inject 10cc per jar. If you don't use SHIPs simply open the lids a crack and squirt in some LI that way.
 
I always make sure the numbers on the syringe are facing me so I can easily see how much liquid is going in each jar.

once all you have finished the last inoculation. take your jars out of the SAB and give each of them a good shake.


If you did everything correctly. Your jars should look like ythis in about 6 days.


That's about it. I hope it works for you.:wave:





Edited by MudaFuka (12/23/15 03:11 PM)


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OfflineBCW
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Re: Liquid Inoculant [Re: MudaFuka]
    #20429805 - 08/16/14 11:03 PM (6 years, 2 months ago)

:takingnotes: Thanks for posting


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OfflinePsiloBeast
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Re: Liquid Inoculant [Re: BCW]
    #20429871 - 08/16/14 11:15 PM (6 years, 2 months ago)

I know we've had our differences Muda...

...but damn, nice Tek

:jokerclap:


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InvisiblePastywhyteM
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Re: Liquid Inoculant [Re: PsiloBeast]
    #20429918 - 08/16/14 11:23 PM (6 years, 2 months ago)

Like the 9er tek but cleaner, I might try this. Nice one muda :rockon:


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Offlinetombosley8
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Re: Liquid Inoculant [Re: Pastywhyte]
    #20430178 - 08/17/14 12:15 AM (6 years, 2 months ago)

just what i've been waiting for and cheaper and easier than I ever thought. Do you think Using a regular mouth qt jar would be ok maybe spread the inoculant out some? And do you think this could be done with a LC solution (2-4%LME) and let colonize some to boost the inoculation power or potency or dilute and make extra inoculant from the one plate. just started using LCs from agar but they take so long to colonise and having to stir is a pain so i guess this idea is much more plausible. I like the no need for colonization aspect just wondering if you think it could help at all... oh yeah how much water do you put in the pc? And how much inoculant per jar? Sorry for blasting you with all that i'm just overly excited and anxious to get it started. Hope that's really not the last grow of yours here this community needs your support (and me too:wink:)


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Re: Liquid Inoculant [Re: tombosley8]
    #20430326 - 08/17/14 12:47 AM (6 years, 2 months ago)

So with this you don't have to wait for a master jar to colonize you can kmock up a whole lot of jars with the same plate!

Very cool


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Invisiblewowimflabbergasted
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Re: Liquid Inoculant [Re: mushpunx]
    #20430331 - 08/17/14 12:49 AM (6 years, 2 months ago)

:fonz:  :congrats:


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OfflineSarah Tonin
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Re: Liquid Inoculant [Re: wowimflabbergasted]
    #20431121 - 08/17/14 04:24 AM (6 years, 2 months ago)

Thanks for sharing the deets, Muda.  Gonna' have to give it a whirl!


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InvisibleImperfect Iam
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Re: Liquid Inoculant [Re: Sarah Tonin]
    #20431151 - 08/17/14 04:37 AM (6 years, 2 months ago)

Great idea Muda, I always wonder when using an agar wedge to inoc. a grain jar with a double wedge, I wonder how far you could stretch this if made into liquid form, I guess this would help give me an idea here, looks like I'm gonna have to burn up another blender making reishi tea, to give me an excuse to buy one that fits jars:wink:


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InvisibleMudaFuka
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Re: Liquid Inoculant [Re: tombosley8]
    #20431203 - 08/17/14 05:08 AM (6 years, 2 months ago)

thanx guys. glad you like it.

Quote:

tombosley8 said:
just what i've been waiting for and cheaper and easier than I ever thought. Do you think Using a regular mouth qt jar would be ok maybe spread the inoculant out some? And do you think this could be done with a LC solution (2-4%LME) and let colonize some to boost the inoculation power or potency or dilute and make extra inoculant from the one plate. just started using LCs from agar but they take so long to colonise and having to stir is a pain so i guess this idea is much more plausible. I like the no need for colonization aspect just wondering if you think it could help at all... oh yeah how much water do you put in the pc? And how much inoculant per jar? Sorry for blasting you with all that i'm just overly excited and anxious to get it started. Hope that's really not the last grow of yours here this community needs your support (and me too;))



im sure a regular mouth qt jar would be fine. that's what I was planning to use originally but all my qts are wide mouth. I'm sure this could be used in conjunction with LC but I think that just raises the chances of contamination. If you want to give your LI a boost just throw in another plate. That's just my opinion so play around with it. See what works best for you.
I don't put much water in the PC since its such a short cycle. Maybe 3/4". As far as how much solution to put in each jar I just try to spread it out evenly threw out the jars I want to inoculate. I've only used this thing twice now so I haven't had time to really play around and get my system down yet. Oh and no that last grow is not going to be my last grow. I took a little bit of time to get my head strait and now I'm back and ready to hit this shit harder than I ever did before.


--------------------
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Edited by MudaFuka (08/17/14 05:38 AM)


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Invisibleblindingleaf
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Re: Liquid Inoculant [Re: MudaFuka]
    #20431505 - 08/17/14 09:02 AM (6 years, 2 months ago)

you want some good shit?? fuck with this TEK!!

awesome dude :thumbup:
im not sure muda could possibly grow more mushrooms (look at his grows) but if he could, this would have to be the only way left :lol:


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Re: Liquid Inoculant [Re: tombosley8]
    #20431542 - 08/17/14 09:15 AM (6 years, 2 months ago)

Quote:

tombosley8 said:
just what i've been waiting for and cheaper and easier than I ever thought. Do you think Using a regular mouth qt jar would be ok maybe spread the inoculant out some? And do you think this could be done with a LC solution (2-4%LME) and let colonize some to boost the inoculation power or potency or dilute and make extra inoculant from the one plate. just started using LCs from agar but they take so long to colonise and having to stir is a pain so i guess this idea is much more plausible. I like the no need for colonization aspect just wondering if you think it could help at all... oh yeah how much water do you put in the pc? And how much inoculant per jar? Sorry for blasting you with all that i'm just overly excited and anxious to get it started. Hope that's really not the last grow of yours here this community needs your support (and me too;))




Wouldn't using LC kind of ruin the whole point of this TEK?  Seeing as how this tek is useful because it WOULDN'T support contams, but if you used LC then you would just be negating the benefits of this tek IMO

Oh, and good job OP. :thumbup:  My agar equipment is arriving on monday,  and this may be one of the first things i try when i have a good isolate/wedge.  Thanks for posting


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Re: Liquid Inoculant [Re: Erlik Khan]
    #20431656 - 08/17/14 10:03 AM (6 years, 2 months ago)

:thumbup:


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Re: Liquid Inoculant [Re: tripdawg420] * 1
    #20431698 - 08/17/14 10:28 AM (6 years, 2 months ago)

not to raid the thread, but just to illustrate that you can use other media than agar slices. all of which use distilled water only.

you can also use this with pf cakes or sliced stem tissue. you use a larger small mouth pint jar and it fits on the end. then you can blend it up and pour it into grain jars. using liquid inoculant.







 


you can make enough liquid inoculant with one pf jar to inoculate 100 quart jars that will colonize in 3 days. its awesome. but generally i don't have that many quart jars lol. its called slurry tek

you can also dump that slurry directly into your bulk sub and colonize a full tub with one pf jar.

 



Oster is another good blender brand that has a detachable blade assembly at the bottom that will fit small mouth mason jars. however, i think the one that mudafuka has found is a little better because the plastic assembly can be pressure cooked. the black plastic assembly on the oster will get warped if it is pressure cooked.
generally when i use this method, ill just clean the black plastic with rubbing alc or spray with lysol. anyway your not touching anything clean on the outside of the jar or the assembly and when your blending its exterior. so as long as you keep your clean work on the inside of the blender jar with clean assembly you shouldn't have any issues. it may look a little ghetto but ive never had a contam in quart jars using this method. id use it more but its too much inoculum and i would be wasting it. if i were running 100 quart jars id probably use this method more often. but this colonizes quart jars in 3 days. after you pour in the liquid amount, you just shake up the jar real evenly. you really only need a small amount per jar for this to work.

ive done a few test runs on 40 quart jars with this method and it colonizes the quarts in 3 days. you just prep all your grain jars and clean work ahead of time. then blend all of it up and pour into jars in one sitting. you would also follow clean technique when doing this.


Edited by eatyualive (08/17/14 11:05 AM)


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Invisibleblindingleaf
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Re: Liquid Inoculant [Re: eatyualive]
    #20431795 - 08/17/14 11:07 AM (6 years, 2 months ago)

That idea is awesome, and way more suitable for bulk substrate inoculation (more nutrients, more noc points), but for grain inoculation, a petri LI I think is more suitable only because plates take way less time to colonize than BRF jars, but to each their own.


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Re: Liquid Inoculant [Re: blindingleaf]
    #20431877 - 08/17/14 11:30 AM (6 years, 2 months ago)

Quote:

blindingleaf said:
That idea is awesome, and way more suitable for bulk substrate inoculation (more nutrients, more noc points), but for grain inoculation, a petri LI I think is more suitable only because plates take way less time to colonize than BRF jars, but to each their own.




true that is quicker. but if you use this in unison with 9er tek its much faster than an agar plate. basically if you cut stem tissue, blend it up in the jar like you see above, suck it up into a syringe, then inject that one 10cc syringe of stem tissue into one pf jar. you can have a fully colonized pf jar stem tissue clone in less than 5 days. i generally get the jar colonized in 3 days using blended stem tissue. then you can blend it up into a quart jar of water and inoculate 100 quart jars and have them colonized in 3 more days. so your talking 100 quart jars fully colonized in less than 10 days starting from stem tissue.

but also one pf jar can become 100 quart jars. if you need 100 quart jars. you can have fully colonized jars in 3 days. im betting one agar plate could inoculate up to 50 but ive not really tried it directly to know the extent of how far you could extend the liquid incoculum


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Edited by eatyualive (08/17/14 11:48 AM)


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InvisibleMudaFuka
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Re: Liquid Inoculant [Re: eatyualive]
    #20431903 - 08/17/14 11:36 AM (6 years, 2 months ago)

My concern with using stem tissue is that it would not be clean enough but I have a shit load of jars, oats are cheep and I like to try as many new things as I can so Ill give it a shot at some point.


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Re: Liquid Inoculant [Re: MudaFuka]
    #20431918 - 08/17/14 11:43 AM (6 years, 2 months ago)

Quote:

MudaFuka said:
My concern with using stem tissue is that it would not be clean enough but I have a shit load of jars, oats are cheep and I like to try as many new things as I can so Ill give it a shot at some point.




also think about this, im injecting it directly into a pf jar and not grain. although i have done it in grain with no issues i don't use as much liquid inoculum in grain jars due to easy contamination.  in grain jars, you have to be very careful about adding too much extra moisture.  if your worried pf jars are pretty damn full proof. the key is to colonize as fast as possible to minimize contamination. this is the reason i use one 10cc syringe per 1 pf jar. at most ill do 2 pf jars per syringe for speed. if your jars done in 3 days. well, thats damn fast. also distilled water only no nutrients added. this may be why the stem tissues stores for years for me with no contamination.  i recently injected some 5 year old clone material and it was so old that it did not colonize but it did not contam.  it simply did nothing.

ive also done the same thing with one 10cc syringe inoculated into 10 pf jars with no contam issues. it takes a little longer to colonize the jars but its still faster than a multispore injection.


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Re: Liquid Inoculant [Re: eatyualive]
    #20431932 - 08/17/14 11:48 AM (6 years, 2 months ago)

I have never made a BRF cake. The closest thing I did to PF tek was a large hemp flower brick. I guess this gives me a reason to give PF tek a shot.


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Re: Liquid Inoculant [Re: MudaFuka]
    #20431941 - 08/17/14 11:50 AM (6 years, 2 months ago)

Quote:

MudaFuka said:
I have never made a BRF cake. The closest thing I did to PF tek was a large hemp flower brick. I guess this gives me a reason to give PF tek a shot.



well, if i were you id use the agar wedges for liquid inoculum. you can probably get away with colonizing more than 20 quart jars of grain per plate. are you planning on sucking up the liquid inoculum into syringes and then injecting it into your grain jars?

then when your bored try out one pf jar. you don't have to be nearly as clean with pf jars.


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