For sterile transfer of mycelium from petri dish to quart jar for expanding liquid culture
Required Equipment
Essential Tools
- Bio safety cabinet/flow hood
- Petri stand
- Petri suction cup
- Scalpel
- Alcohol tray
- Bacti-Incinerator
- Parafilm
- Labels
Safety & Sanitation
- 70% Isopropyl alcohol
- Sterile arm sleeves
- Nitrile gloves
- Alcohol
- Towelettes
Cultures And Prepared Media
- Source petri dishes
- Sterilized liquid culture jars (Pre-prepared with nutrient broth)
Pre-Transfer Procedure
Initial Setup
- Turn on and preheat
- Bacti-Incinerator
- Wipe base with alcohol
- Position in hood
Bio Cabinet Preparation
- Don gloves and arm sleeves
- Sanitize hood interior and arm ports
- Lower hood sash
- Arrange workspace left-to-right
Equipment Sterilization
- Sanitize all equipment with alcohol
- Carefully clean petri dishes (avoid label smearing)
- Place materials in hood following directional workflow
Transfer Procedure
Workspace Setup
- Adjust chair height for optimal reach
- Sanitize gloves thoroughly
- Remove parafilm from source petri
- Position dish face-down on stand
- Pre-loosen jar lids under laminar flow
Sample Collection
- Attach suction cup to working petri
- Sterilize scalpel (25 seconds in Bacti-Incinerator)
- Cool scalpel in air flow
- Lift petri with suction cup facing upward
- Cut triangular sample from outer colony edge
- Collect sample on scalpel tip
Jar Inoculation
- Return petri to stand
- Remove jar lid maintaining sterile technique
- Transfer sample into broth
- Tap scalpel gently to release sample
- Reseal jar ensuring proper gasket alignment
Post-Transfer Protocol
Finishing Steps
- Reseal petri dish with fresh parafilm
- Label jars with: Species
- Date Source plate info
- Place jars in incubation room
- Return source plates to storage
Quality Control Notes
- Maintain sterile technique throughout
- Work within laminar flow Avoid crossing over sterile field
- Monitor for contamination signs
- Document all transfers