Liquid Inoculant ( NEW version)

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MudaFuka
Poppin bottles

Registered: 12/14/13
Posts: 18,648
Loc: Canada

Liquid Inoculant ( NEW version)

40
#20429745 - 08/16/14 08:53 PM (12 years, 4 days 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 01:11 PM)

Extras:

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