Preparation of Bacterial Inoculum for Acinetobacter Biofilm Formation (Video) | JoVE
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Bacterial Inoculum Preparation For Acinetobacter Biofilm Formation:
1. Inoculate A Blood Agar Plate With 2-10 μL Of The Bacterial Strain Using A Sterile Pipette Tip. Streak The Agar SurfaceThinning It Out With Intermittent Flaming. Incubate The Plate At 30°C For 20-24 Hours.
2. Select A Single Colony From The Agar Plate And Inoculate 5 ML Of Sterile BHI Broth. Incubate The Inoculated Broth In A Shaking Incubator At 30°C150 Rpm For 20-24 Hours.
3. Transfer 1 ML Of The Culture To A Sterile Microcentrifuge Tube And Centrifuge At 6000 G For 10 Minutes At Room Temperature. Discard The Supernatant.
4. Add 1 ML Of Sterile Phosphate-buffered Saline (PBS) To The Pellet And Resuspend Using A Vortex. Centrifuge At 6000 G For 10 Minutes At Room Temperature And Discard The Supernatant.
5. Resuspend The Resulting Pellet In Sterile10-fold Diluted BHI Broth Using A Vortex. Ensure The Optical Density Is Around 0.1 At 600 Nm
Cite This Article
Preparation of Bacterial Inoculum for Acinetobacter Biofilm Formation. J. Vis. Exp. (Pending Publication), e200399, doi:10.3791/200399 (2023).