Mushroom Liquid Culture Recipe

Materials Needed

• 500 mL distilled water

• 5–7 g honey or 5–7 g glucose syrup

• 1 500 mL mason jar with:

  • Self-healing injection port

  • Breathable filter (micropore tape, synthetic filter disc, or polyfill)

• 1 sterile syringe (10–20 mL)

• Mycelium (agar wedge or clean grain spawn) or spore syringe

• Pressure cooker or autoclave

• Isopropyl alcohol (70%+)

• Gloves, face mask, paper towels

• Laminar flow hood or still air box (for sterile work)

🧑‍🍳 Step-by-Step Instructions

1. Prepare the Nutrient Broth

• In a clean container, mix:

  • 5–7 g honey or glucose syrup

  • Into 500 mL distilled water

• Stir well until completely dissolved.

🔬 Tip: For best results, aim for a sugar concentration of 1–2% by weight.
(5 g in 500 mL = 1%, 7 g = 1.4%)

2. Fill the Jar

• Pour the liquid into your mason jar, filling to about 350–400 mL to leave headspace.

• Secure the lid with:

  • Injection port

  • Filter for gas exchange

3. Sterilize

• Place the jar in a pressure cooker.

• Sterilize at 15 PSI for 20–30 minutes.

• Let cool to room temperature before inoculating (minimum 8 hours or overnight).

4. Inoculate

• Wipe all surfaces, gloves, and tools with isopropyl alcohol.

• Use a sterile syringe to inject:

  • 1–2 mL of spore solution or liquid culture

  • OR drop in a small agar wedge (in sterile conditions)

• Shake gently to distribute.

5. Incubate

• Store at appropriate temperature:

  • Lion’s Mane: 21–24°C (70–75°F)

  • Cordyceps militaris: 18–20°C (64–68°F)

  • Oysters: 21–24°C (70–75°F)

• Keep in the dark or low light.

• Shake every 3–4 days once growth is visible.

6. Check for Growth

• Within 3–10 days, look for:

  • Cloudiness or white wispy strands (healthy mycelium)

  • Avoid colorful growth (contamination)

7. Use or Store

• When fully colonized, extract with sterile syringe.

• Store in refrigerator (2–8°C) for up to 6 months.