Materials: 

2 cell culture conical tubes

95% Etoh

Inoculation Loop

Bunsen Burner

Striker

Gloves

10mL LB Broth (ATCC 15224) 

Escherechia coli (Prepared 10/01/2012 by CM)

5mL Pipette

Pipette Aid



Method: 



1. Lit burner and adjusted flame. 
2. Opened pipette and attached to pipette aid aseptically.
3. Flamed source bottle of LB Broth and transferred 5mL  LB broth into each conical tube. 
4. Disposed of pipette in biohazard receptacle. 
5. Opened inoculation loop and rinsed in 95% Etoh then sterilized loop using burner flame. 
6. Aseptically transferred approximately 1 colony from ATCC E. coli plate into each broth tube re-sterilizing loop between transfers. 
7. Flamed broth source bottle and returned to bench shelf. 
8. Flamed loop again to sterilize, returned to storage container.
9. Extinguished flame and returned burner and striker to their storage areas. 
10. Labeled culture and put in 37 degree C shaking incubator which delivers aeration to help grow and provide constant nutrient supply.
11. Wiped down bench with disinfectant. 



Data and Results: 

10/02/2012 TBD after 24 hours. 

10/03/2012 Inoculation successful. 



Discussion:

Will use the growth in broth to inoculate agar plate next. Busted myself not tilting broth source during first 5mL transfer to the tube. Will be more mindful of that going forward. 



Conclusion: 

From visual inspection of tubes I can see colony in Tube 2 floating on top of broth. Inoculating is fun!