CULTIVATION GUIDE

Cultivation Guide

This guide outlines the core stages of the cultivation process, from initial setup through to harvest and post-harvest handling.

Each step builds on the previous, with a focus on maintaining clean technique, stable environmental conditions, and consistent workflow.

While individual methods may vary, the principles remain the same: control contamination, support healthy mycelial growth, and create conditions that allow for reliable development.

Mycelial growth under sterile conditions.

A small number of foundational tools and materials are required to create a clean and controlled environment for cultivation.

Many cultivators begin with a still air box (SAB) or similar enclosed workspace to reduce airborne contamination during handling. For more advanced setups, a laminar flow hood provides a higher level of control and consistency.

Proper preparation of materials is essential. Substrates and grain are typically sterilized or pasteurized using a pressure cooker to minimize competing microorganisms prior to inoculation.

Cultures may be introduced using spore syringes or liquid cultures, depending on preference and experience. As techniques develop, many cultivators transition to working with agar to refine and stabilize cultures before expansion.

As the process progresses, consistency in technique and environmental control becomes increasingly important. Each stage builds on the previous, with small variations in conditions directly influencing overall results.

STEP 1 - PREPARING SPAWN MEDIUM

Once a sterile workspace is prepared, the next step is selecting and preparing your spawn medium. Rye grain is commonly used for its consistency, moisture retention, and strong colonization performance.

Organic, NON-Fungicide Rye Grain Berries

1. Rinse

Place the grain into a large container or 5 gallon bucket and rinse with cold water, pouring off the water several times until it runs noticeably clearer.

This removes dust, debris, and excess starch that can contribute to contamination.

2. Soak (24 hrs)

Cover grain with water and allow it to soak for approximately 24hrs.

This fully hydrates the grain and activates dormant endospores, making them easier to eliminate during sterilization.

3. Heat Treatment (20 minutes)

After soaking, drain the grain and refill with hot water (not boiling). Let it sit for ~20 minutes.

This allows the grain to absorb additional moisture while maintaining its structure.

4. Drain & Dry

Strain the grain thoroughly using a colander and allow it to air dry until the exterior is no longer wet.

Surface moisture should be minimal to reduce the risk of bacterial growth during colonization.

Simple test: a few grains may stick to your hand, but they should not feel wet or leave residue.

Rye grain drying on a mesh screen under controlled lab condition.

5. Load & Seal

Transfer the prepared grain into your chosen containers.

Jars: Fill to an appropriate level (~ 3/4 full) to allow for proper air flow and colonization. Use heat-resistant glass jars suitable for pressure sterilization. Ensure all lids are properly tightened and fitted before sterilization.

Filter patch bags: Fill (~1/2 full), remove excess air, and seal using heat sealer to create secure, sterile closure.

Proper sealing is critical —this step helps maintain sterility and prevents contamination during the sterilization and incubation process. All containers must be able to withstand high heat and pressure during sterilization.

6. Sterilization

Place containers or bags into a pressure cooker:

Stovetop - 15 PSI for ~2.5 hrs

Electric - 15 PSI for ~4 hrs

If your sterilizer does not have to capacity to pressurize to 15 PSI, add 1 hr to cook time.

Container NOTES:

If using mason jars, lids with an integrated inoculation port are recommended for controlled inoculation.

For bags, use autoclave filter patch bags (Type 3T, 0.2 micron). Prior to sterilization, create a small slit in the corner of bag to allow for pressure equalization and prevent bag rupture.

After sterilization, re-seal immediately using heat sealer or mushroom bag clamp.

7. Cool & Rest

After sterilization, allow pressure to return to zero naturally before opening sterilizer.

Before opening, wipe down the exterior surface and lid of the sterilizer with a clean cloth or disinfectant.

Once pressure has fully released, let the containers remain inside the sterilizer and cool undisturbed for 8-12 hrs (or overnight).

Avoid handling or moving containers or bags while hot, as this can introduce contamination through pressure differentials or compromise seals.

Slow, undisturbed cooling helps maintain sterility and prepares the grain for clean inoculation.

STEP 2 - INOCULATION

Once your containers have fully cooled and been removed from the sterilizer, they are ready for inoculation.

Inoculation should be performed within 24 hours of sterilization. Grain contains a finite amount of moisture that must support the entire colonization process. Delaying inoculation can reduce potential yield and create a less optimal environment for mycelial growth.

Liquid culture introduced into grain using sterile technique in a controlled workspace.

1. Prepare Workspace

Set up in front of a laminar flow hood or inside a still air box (SAB). Thoroughly disinfect all surfaces, tools, and gloves before beginning.

This stage carries the highest contamination risk —clean technique directly determines success.

2. Prepare Materials

Gather your fully cooled grain containers, liquid culture syringe, sterile 18-gauge needle, alcohol or disinfectant, and paper towel or wipes.

Having everything within reach minimizes movement and exposure during inoculation.

3. Sanitize Surfaces & Syringe

Wipe down the exterior of the syringe, containers, your gloves and forearms. Re-sanitize hands frequently throughout the process.

Reducing surface contamination lowers the chance of introducing competing organisms.

4. Attach & Sterilize Needle

Attach a sterile 18-gauge needle (included with liquid culture syringe products) to your syringe. Sterilize needle using flame (preferred) or disinfectant before use.

Sterilizing the needle ensures a clean entry point into the container.

5. Inoculate the Container

Insert the needle into the inoculation point:

Through an injection port lid, if using jars with ports
Through a pre-drilled hole (covered with micropore tape after injection)
Or directly into a filter patch bag

Inject approximately 5 mL of solution per 1 L of container volume. Using slightly more solution can accelerate colonization; however, excessive moisture should be avoided.

Proper distribution supports faster and more uniform colonization.

6. Distribute Liquid Culture

Gently shake or rotate the container immediately after injection to disperse the culture.

This creates multiple inoculation points, improving colonization speed and consistency.

7. Seal & Secure

Ensure injection ports reseal, lids are tightened, or bags are properly sealed (heat sealer or clamp if applicable)

Maintaining a closed, sterile environment is critical after inoculation.

8. Transfer to Colonization Area

Move containers to a clean, stable environment with minimal airflow and disturbance.

Limiting movement and exposure allows mycelium to establish without competition.

9. Work Efficiently

Avoid unnecessary exposure of sterile materials and complete the process smoothly and deliberately.

Short exposure time reduces contamination risk.

LC Best Practice NOTE:

Many cultivators use one full 10mL syringe per container (standard jars or small-mid bags) to promote faster, more aggressive colonization, though lower volumes can still be effective.

STEP 3 - COLONIZATION

Colonization is the period where mycelium spreads throughout the grain, building a strong, healthy foundation for fruiting.

Fully colonized grain, showing dense and even mycelial growth.

1. Distribute Inoculant

Immediately after inoculation, gently shake or rotate the container to distribute the liquid culture and create multiple inoculation points.

This supports faster and more uniform colonization.

2. Allow Initial Growth

Let the mycelium establish for 24-48 hrs without shaking or disturbing.

Give the mycelium time to take hold and begin spreading.

3. Store in a Clean, Stable Environment

Place containers in a clean area with minimal airflow and disturbance.

A stable environment reduces contamination risk and promotes consistent growth.

4. Maintain Ideal Temperature

Maintain a consistent temperature between 21–24°C (70–75°F) for optimal colonization.

Below 21°C (70°F) slows growth; above 24-25°C (75-77°F) may accelerate colonization, but can increase contamination risk and cause heat buildup within the grain.

It is crucial to keep temperature below 26.5°C (80°F) throughout colonization.

5. Humidity Is Not Crucial

Containers are sealed, so ambient humidity is not important.

Normal indoor conditions are ideal. Avoid excessively damp environments.

6. Avoid Air Flow and Handling

Keep containers away from fans, vents, or drafts. Minimize movement and handling, especially during the first stages.

Stability is key to strong, uninterrupted colonization.

7. No Light Required

Keep containers in ambient or low-light conditions.

Mycelium does not need light. Complete darkness is not necessary.

8. Monitor Progress

Mycelium will spread throughout the grain, turning it white and forming a dense, uniform network.

9. Typical Timelines

Spore syringe: ~2-6 weeks

Liquid culture: ~2-4 weeks (typically faster due to active mycelium)

Faster colonization is associated with proper inoculant distribution, stable temperatures, and optimal grain moisture. Slower growth may indicate suboptimal conditions but does not necessarily mean failure.

10. OPTIONAL (20-30% Colonization)

Gently shake or break up the grain to redistribute colonized material and accelerate overall colonization.

Consistency in temperature, cleanliness, and handling directly influences colonization speed and culture strength.

11. Signs of Full Colonization

Grain is barely visible
Mycelium is bright white and uniform
No uncolonized or discoloured areas remain
The structure appears dense and fully connected

Patience and consistency during this stage are critical— stable conditions will always outperform constant adjustment. Fully colonized grain should appear uniform and stable before proceeding to the next stage.

SL- Final steps after pasteurization.PNG

STEP 5 - MIXING SPAWN WITH SUBSTRATE

This step involves combining fully colonized grain spawn with prepared substrate, a process often referred to as spawning to bulk.

1. Prepare Your Workspace

Work in a clean, low-traffic environment.

Before beginning:

Wipe down all surfaces
Minimize airflow (avoid fans, drafts, vents)
Wear gloves if possible

Full sterile conditions are not required, but clean technique helps reduce contamination risk.

2. Prepare Your Fruiting Container

Use a clean tub, tray, or tote.

Ensure the container is:

Wiped down prior to use
Dry and free of debris

Place a clean garbage bag inside the container to act as a liner, pressing it into the corners so it sits flush with the tray.

Liners help maintain consistent moisture, reduce side pinning and allow the substrate to contract naturally during colonization.

3. Add Substrate and Grain Spawn

Open your cooled, pasteurized substrate and fully colonized grain spawn.

Add both directly into container.

We recommend a ratio of:

~25% grain spawn / 75% substrate

4. Break Up & Distribute Spawn

Break apart the colonized grain and distribute it evenly throughout the substrate.

Ensure:

No large clumps remain
Spawn is evenly spread

This creates multiple inoculation points and supports faster colonization.

5. Mix Thoroughly

Combine the materials until the mixture is consistent throughout.

The goal is even distribution across the entire substrate.

6. Level the Surface

Lightly level and gently compress the surface to create an even substrate layer.

Using a flat tool (such as a large BBQ flipper or spatula) can help achieve a smooth, consistent finish.

This helps:

Create an even colonization environment
Prevent low spots where moisture can collect
Reduce the risk of pooling during incubation

The surface should be:

Flat
Evenly distributed
Lightly compacted (not densely packed)

7. Seal & Label

Place the lid on the container, cut the garbage bag about 2” below the lid all the way around the tray, and remove the excess bag.

Using a permanent marker, label the container with:

Date
Strain
Any relevant notes

8. Ready for Bulk Colonization

The substrate is now ready to colonize.

Place the container in a clean, stable environment and allow the mycelium to fully colonize before introducing fruiting conditions.

Even mixing, proper ratios, and clean handling at this stage directly influence colonization speed, uniformity, and overall yield.

STEP 6 - INCUBATION (BULK COLONIZATION)

Once the substrate has been mixed and the container sealed, it enters the incubation phase, where mycelium spreads throughout the substrate and fully establishes.

This stage typically takes :

~7-10 days (May vary depending on conditions and genetics)

1. Maintain Stable Conditions

Place the container in a clean, low-traffic area, and avoid disturbance.

Do not open the container
Avoid moving or handling unnecessarily
Minimize airflow (no fans, drafts, vents)

Consistency during this stage is critical for even colonization.

2. Temperature Control

Maintain temperature within: 21–25°C (70–77°F)

Avoid exceeding : 27°C (80°F)

Higher temperatures can lead to contamination or overheating within the substrate.

3. Lighting

Light is not required during incubation.

Ambient light is fine
Complete darkness is not necessary

4. Monitor Moisture

Check periodically for:

Condensation buildup
Water pooling on the surface

If needed:

Briefly remove lid
Wipe excess moisture
Replace lid and maintain stable conditions

5. Observe Colonization

Healthy growth appears:

Bright white
Evenly spreading
Dense and connected

The surface will gradually become fully covered.

6. Watch for Contamination

Remove immediately if you observe:

Green, black, grey, or pink growth
Wet/slimy areas
Strong or unusual colours

Early removal helps prevent spread.

STEP 7 - TRIGGERING FRUITING

Fully Colonized

The substrate is fully incubated when:

Surface is completely white
No uncolonized areas remain
Growth is uniform and stable

Allow an additional 1-2 days after full coverage before introducing fruiting conditions.

Stable conditions will always outperform constant adjustment.

Once the substrate is fully colonized, fruiting is initiated by adjusting environmental conditions.

This transition signals the mycelium to shift from colonization into fruit body formation.

Key Environmental Changes

To trigger fruiting, adjust the following:

1. Humidity

Increase humidity to support pinning and fruit development.

Maintain high humidity (typically 85-95%)
Moisture availability is critical, as fruiting bodies are composed of ~90% water

2. Fresh Air Exchange (FAE)

Increase airflow to introduce oxygen and reduce CO₂ buildup.

Move from low-airflow incubation → active air exchange
Proper airflow helps prevent elongated or deformed growth

3. Light Exposure

Introduce a consistent light cycle.

12 hrs light / 12 hrs dark
Ideal spectrum: 6000-7000K (cool white)
Light acts as a directional and developmental trigger (not a primary energy source)

Why This Works
These changes mimic natural conditions.

In nature, mycelium grows in dark , low-oxygen environments (like soil or compost).

As it reaches the surface, it encounters:

Light
Fresh air
Increased moisture

These signals trigger the formation of fruiting bodies.

Final Transition

Shift from dark, low-airflow environment → controlled fruiting conditions
Maintain consistent humidity, airflow and lighting
Avoid frequent environmental fluctuations

Fruiting is triggered by environmental change —consistency after the shift determines quality, uniformity, and overall yield.

STEP 8- FRUITING CONDITIONS & MAINTENANCE

1. Introduce a Fruiting Dome
At this stage, replace the lid with a fruiting dome to increase airflow while maintaining humidity.

Use the same container/tub/tote used to build the fruiting substrate, flipped upside down
Place it directly over the tray to act as a dome
Cut/drill small holes (~2") in the dome to allow passive airflow
Cover holes with breathable filter material (ie:// polyfill)
Secure dome to bottom container with clips if needed to maintain positioning

This setup creates a controlled environment that balances fresh air exchange with humidity retention.

2. Increase Humidity
Humidity should be raised to support pin formation and early fruiting.

Lightly mist the environment as needed
Mist when there is no visible condensation on the dome walls
Use a fine mist setting —avoid large droplets

The goal is a lightly hydrated surface, not pooling water.

3. Adjust Misting Based on Conditions
Misting frequency will vary depending on your environment.

High ambient humidity → Mist less frequently
Dry environments or increased airflow → Mist more often

You can skip misting if:

Condensation is already present on dome walls
Surface moisture is sufficient
Water pooling is observed

If excess moisture builds up:

Remove the lid briefly
Wipe condensation from the dome
Allow slight air exchange before resealing

4. Balance Humidity & Airflow

Optimal fruiting requires balancing both:

Maintain high humidity within the dome
Allow consistent fresh air exchange
Avoid overly stagnant or overly dry conditions

Aim for the highest humidity possible while still allowing clean airflow through the dome.

Airflow NOTE:

Ambient room airflow is beneficial
Avoid directing fans at the substrate or dome
If using a fan, keep it indirect to gently circulate air in the room.

Direct airflow can dry the substrate and disrupt surface conditions.

5. Pin Formation (Primordial Stage)

After approximately 7-10 days, small pins will begin to form.

These will rapidly develop into fruiting bodies within 5-7 days.

At this stage:

STOP misting the substrate surface directly
Maintain humidity by misting the sides of the dome instead

Surface conditions drive pinning. Stable humidity and airflow are key to consistent development.

Primordia (pins) beginning to form.

STEP 9 - HARVESTING

Harvesting marks the transition from cultivation to collection —where your hard work begins to yield results.

Mushrooms are ready to harvest when the veil beneath the cap breaks and the cap begins to open. Timing is important —harvesting at this stage preserves structure, cleanliness, and overall quality.

1. Identify the Right Time to Harvest

Harvest just after the veil breaks
Caps should be slightly open, not fully flattened
Earlier harvest = cleaner fruits and reduced spore drop

2. Harvest Cleanly

Use clean, sharp scissors or snips to cut at the base of the stem.
Avoid pulling to prevent disturbing the substrate
Handle mushrooms minimally to protect the surface

Clean technique helps preserve future flushes.

3. Continue Harvesting Through the Flush

Harvest daily as mushrooms mature at different rates
Once a full flush is complete, lightly mist the substrate surface
Maintain humidity to support additional flushes

Multiple flushes can occur until nutrients are depleted.

4. Expected Yield Cycles

Typical grows produce 1-2 strong flushes
In controlled environments, up to 5-6 flushes may occur
Dispose of substrate once contamination appears or yield declines

5. Taking Spore Prints (Optional)

For genetic preservation or future work:

Allow cap to open approximately halfway
Remove the cap and place it gill-side down on sterile foil
Leave for ~12 hrs in a clean, still-air environment
Remove the cap from the foil and allow the print to dry
Scrape the spores into a sterile aqueous solution (sterilized water) and draw the solution into a syringe to create a spore syringe!
Or keep the spore print for long term storage (prints remain viable for up to 10 years, possibly longer)

Consistent harvesting technique, clean handling, and proper timing directly influence yield quality, longevity of flushes, and genetic preservation.

STEP 10 - DRYING

Proper drying is essential to preserve quality and prevent contamination. Mushrooms must be fully dried before storage.

1. Prepare for Drying

After harvesting, gently remove any excess substrate. Avoid washing with water, as added moisture will slow the drying process.

2. Arrange for Airflow

Place mushrooms in a single layer on a drying rack, screen, tray or in dehydrator. Ensure space between them to allow consistent airflow.

3. Dry Using Controlled Conditions

Dehydrator (Recommended): Dry at 35-45°C (95-115°F) for 6-12 hrs depending on size
Air Drying: Place in a low-humidity, well-ventilated space for 2-4 days. A fan can be used to gently circulate air, but avoid direct, strong airflow.

4. Check for Full Dryness

Mushrooms are fully dry when stems snap cleanly rather than bend. Any remaining moisture can lead to spoilage.