1. LAB SAFETY

2. Apparel Shoes Lab coats Safety glasses

3. Note! No eating, drinking, or smoking in the lab at any time.

4. Spills Wash Notify Receive instructions

5. Accidents Be careful! Pay attention Report any injuries

6. Other Matters Broken glass Other waste Pipetting Volatile chemicals Dirty labware Labeling General cleanliness

7. ASEPTIC TECHNIQUE

8. I. Contaminants A. Bacteria, fungi, and insects B. Initial contaminants C. Latent or persistent contamination D. Introduced contamination E. Detection of contaminants

9. THE TRANSFER HOOD The hood should remain on –Necessary items present, unnecessary things removed Check the bottom of the hood for blockage Wash hands

12. THE TRANSFER HOOD Spray or wipe the inside of the transfer hood Wipe hands and lower arms with 70% EtOH Remove dangling sleeves and jewelry

13. Sterilization and use of supplies and equipment Know which of your implements, flasks, etc. are sterile and which are not Monitor autoclave time Sterilized items should be used within a short time Items that come packaged sterile should be examined carefully for damage before use

14. Sterilizing tools, vessels etc. Autoclaving Ethylene oxide gas UV radiation Dry heat Microwave

15. Working in the transfer hood: Work well back in the transfer hood Make sure that materials in use are to the side of your work area, so that airflow from the hood is not blocked Instruments (scalpels, forceps) can be sterilized by flaming or immersion in hot beads

16. Working in the transfer hood: Plant material should be placed on a sterile surface when manipulating it in the hood Remove items from the hood as soon as they are no longer needed When transferring plant cultures, do contaminated cultures last.

17. Working in the transfer hood: When finished in the hood, clean up after yourself. Be sure when you are finished that you turn off the gas to the burner It is pointless to practice good sterile technique in a dirty lab

18. Surface-sterilizing plant material Care of stock plants Alcohol Bleach Calcium hypochlorite Mercuric chloride Hydrogen peroxide Rinsing

19. In the media Use of antibiotics and fungicides Plant Preservative Mixture Rinsing