MUSHROOM

LAB 101
Rod Stafford
August 2014

WHAT CAN YOU GROW AT

HOME?
Start with something easy

any oyster variety (white, grey, blue, yellow, pink,

brown)

shiitake (albinos)
reishi (antlers and conks)
black poplar (exotic?)
turkey tail (rosette forms)
Panellus stipticus (bioluminescent)
hen of the woods, chicken of the woods,
lions mane, bears head take a little more
time

Inoculate dowels for log cultivation

GOALS
1. General Cultivation Overview
2. Making a Glove Box (Old-Tech)
3. Working with Agar
4. Producing Grain Spawn

NOT COVERED
Spawn Techniques (other than grain)
Commercial, Large-Scale or Bulk Growing
H2O2 Techniques
Laminar Flow Technique
Strain Isolation
Anything Illegal

CHEATING
MOTHER NATURE
Even though a
mushroom may
produce millions of
spores, complex
interactions among
hundreds of other
fungi, bacteria, etc.
in nature maintain
an ecological
equilibrium.

Source: Growing Gourmet and Medicinal

Mushrooms, Stamets

For
mushrooms to
grow free from
competition,
provide an
environment as
sterile as a
semi-conductor
manufacturing
factory.

SOURCE MATERIAL
1. Spores
2. Mycelium
A. Buy
i.
ii.

Syringe
Agar Plate

B. Library
i.
ii.

Agar Slant/Plate
Cardboard Pellets

3. Living Tissue

PARTS LIST TO MAKE RODS

GLOVE BOX
A transparent storage bin (Walmart, Target, etc) with a lid,

24L x 16W x 12H

2 fluorescent light fixtures 120V (IKEA, etc.)
velcro tape (Hancock Fabrics, Michaels, etc)
254 nm uv germicidal bulb, 8 Watt G8T5 (11.3)
(uvclamps.com, atlantalightbulbs.com, etc) ($6.39)
Hole saw (Harbor Freight, etc)
or Drill bit and hack saw (any hardware store)
Disposable Tyvek suit (Amazon, Lowes, etc.)

Total cost about $25 - $75

SAFETY ISSUES
Never expose bare skin under a germicidal lamp
Do not look at a germicidal lamp without protection
Do not use an open flame inside a glove box

ANOTHER OPTION
makezine.com/projects/home-mycology-lab/

PARTS LIST
GLOVE BOX WORK ACCESSORIES

Disposable Gloves
Isopropyl alcohol
Disinfectant spray/wipes
Parafilm*

ITEMS DOWN THE ROAD

Scalpel (handle and blades)

Inoculant Loop
Filter Disks for mason jars
Micropore Tape
Self-Healing Injection Ports
Empty syringes (BD Luer-Lok) w/ needles (16G)

Working with Agar

Agar is an extract from a seaweed used to solidify media

Functions as a scaffolding, it has no nutritive value

Allows you to observe growth in two dimensions

Petri Dishes

A petri dish is a shallow cylindrical glass or plastic lidded dish

that biologists use to culture cells
My preferred specs for mushroom culturing:

Disposable plastic
100 x 15 mm
Gamma sterilized
Solid vs segmented

WORKING WITH AGAR

Make your own
Pros

Cons

Make as many as you need

Cheaper per plate

Requires sterile environment

More prone to contamination

Purchase pre-made
Pros

Cons

Convenience
Uniform, sterile plates

More expensive
Usually in batches of 10-20

WORKING WITH AGAR

Agar Sources (there are many more)

mycosupply.com
Amazon
EBay
fungiperfecti.com
sciencebobstore.com/products/Nutrient-Agar-Powder.html

WORKING WITH AGAR

Types of Agar Formulations
General Purpose Fungi
Potato Dextrose (PDA)
Malt Extract (MEA)
Cornmeal
1% Peptone Agar
Dog Food/Rabbit Food/Ovaltine.etc.
Yeasts and Molds
Brain Heart Infusion Agar (BHIA) with 5% Sheep Blood
Specialty
Czapek Dox Agar (for Penicillium)
Sabouraud's Dextrose (dermatophytes)
Tryptic Soy Broth (for bacteria, not fungi!)
many others..

WORKING WITH AGAR

Agar Notes
25 grams of agar will make ~ 30 plates
Buy pre-mixed agar to save time and effort!
Sterilize MEA for 35 minutes at 15 psi!
Provide gas exchange during sterilization!

I use a 500ml glass liquor bottle with a plastic cap, drill a hole in the cap
and use a square of Tyvek

WORKING WITH AGAR

To make your own plates you will need:
A.
B.
C.
D.
E.
F.
G.
H.
I.

Sterile Petri Dishes

Nutrient Agar
Distilled Water
Pressure Cooker
Bottle to sterilize solution
Graduated Cylinder and/or Scale
Piece of Tyvek or cut a filter disc
Aluminum foil
Glove Box and GB goodies

breathable foam plugs

WORKING WITH AGAR

1. Dissolve agar in water at a ratio of 1:20
25 g MEA in 500 ml H2O for ~ 20-30 plates
5 g MEA in 100 ml H2O for ~ 6 plates

2. Cover flask with aluminum foil and sterilize at 15

P.S.I for 15 minutes in pressure cooker (avoid
caramelizing sugars)
3. Remove from PC once pressure normalizes
4. Allow to cool to ~ 150F and place in glove box
5. In glove box, stack plates 3-6/stack
6. Working from bottom to top, pour plates ~ 1/3 full
7. Allow to cool while stacked

GRAIN SPAWN
Rye Berries, aka winter wheat grain
larger kernel size helps prevent clumping
commonly available
low in bacterial endospores

GRAIN SPAWN PREPARATION

add CaCO3 and Ca2SO4
soak raw grains overnight - forces
germination of endospores
bring to boil for 10 minutes

GRAIN SPAWN PREPARATION

drain in colander

use steam from

cooking to dry berries

GRAIN SPAWN PREPARATION

fill quart canning jars
1/3 to 1/2 full with
grain

GRAIN SPAWN PREPARATION

use filter disks or

tyvek squares under

lids
cover with aluminum
foil

GRAIN SPAWN PREPARATION

sterilize for 90 minutes at 15 p.s.i.

GRAIN SPAWN PREPARATION

allow to cool COMPLETELY before
using, store covered until use

AGAR TO GRAIN TRANSFER

Place all the grain jars in your GB along with the agar
culture

Loosen the cap from each jar but dont remove them
Flame sterilize your scalpel and place it in the glove
box to cool

or use a pre-sterilized blade inside the GB

Open the culture dish and cut a small wedge of agar

Spear the wedge with the scalpel blade, replace the
culture lid, lift a lid from a grain jar and drop the
wedge inside without touching anything

repeat with 1 or 2 more wedges before moving to next

jar

SUCCESS

MAKING DOWELS IS EASY

RESOURCES
WEBSITES
makezine.com/projects/home-mycology-lab/
www.mykoweb.com/articles/cultivation.html
shroomery.org
SPORES/MYCELIUM/SPAWN
ebay (Caveat emptor)
sporeworks.com
fungiperfecti.com
plantpath.psu.edu/facilities/mushroom/cultures-spawn
(very pricey, but they have EVERYTHING)

SUPPLIES
www.mycosupply.com
www.cynmar.com
www.mycopath.com/