Hello everyone,

I'm quite new to culturing anaerobic bacteria. I'm feeling both nervous and excited about this new endeavor!

Our lab operates on a limited budget, and we currently have access to an anaerobic jar (with gas packs and indicators), sterile loops, Petri plates, a 37°C incubator, a standard refrigerator, and -20°C and -80°C freezers.

When I receive the purchased F. nucleatum strain, what are the crucial first steps I need to take to successfully revive and grow it in our lab setting?

Specifically, I would be very grateful for advice on:

1. Initial Resuscitation: What is a recommended, cost-effective protocol for resuscitating the culture upon arrival?

2. Essential Media: Which growth media (including specific components if possible) would you suggest for consistent and cost-effective growth and maintenance of this strain?

3. Anaerobic Jar Usage: Are there any specific tips or tricks for successfully culturing F. nucleatum using only an anaerobic jar?

4. Basic Required Reagents/Supplies: Beyond the standard lab equipment I've mentioned, what are the essential reagents or supplies I absolutely need to have on hand to work with this bacterium?

5. Short-Term and Long-Term Storage: What are cost-effective methods for both short-term maintenance and long-term preservation of F. nucleatum cultures to ensure viability?

Any detailed guidance, even on seemingly small steps, would be incredibly helpful for someone like me who is just starting with anaerobic bacteria. I'm eager to learn and successfully work with this fascinating organism!

Thank you in advance for your time and expertise.

Bilal