Required Materials
- Flow hood or glovebox
- 70% isopropyl alcohol
- Paper towels
- Nitrile gloves
- Mask
- Scotch tape
- Liquid culture syringe
- Sterilized grain bags
Pre-Inoculation Setup
Work Area Preparation
- Turn on flow hood (if using)
- Create a clean workspace
- Gather all materials within reach
Personal Protection
- Put on nitrile gloves and mask
- Thoroughly sanitize gloves with 70% alcohol
- Ensure coverage of entire glove surface, including between fingers
Inoculation Procedure
Prepare Sanitizing Pad
- Saturate paper towel with 70% alcohol
- Ensure towel is wet but not dripping
Wipe Injection Site
- Thoroughly wipe injection area on grain bag
- Avoid contact with white filter patch
- Allow alcohol to evaporate briefly
Culture Preparation
- Vigorously shake culture syringe for 30 seconds
- This distributes mycelium cells evenly
Needle Setup
- Carefully open sterile needle package
- Remove syringe cap
- Attach needle firmly to syringe
- Remove needle cap
Inoculation Process
- Insert needle through prepped injection site
- Inject exactly 1ml of liquid culture
- Withdraw needle smoothly
Post-Injection
Sealing
- Cut fresh 1-inch piece of tape
- Ensure tape hasn't been previously exposed
- Seal injection point completely
Final Steps
- Unroll grain bag if rolled
- Gently mix contents to distribute culture
- Store in incubation
Important Notes
- Never inject through the filter patch
- Maintain sterile technique throughout
- Work efficiently but carefully
- Document date and culture strain used