Hello everyone! I have been trying to develop a protocol for my thesis research which includes culturing and storing Bifidobacteria. I have come across many articles and studies related to the topic but non clearly explains the specific details of how to prepare the media, specifically with respect to adding L-cysteine HCl to this media (Agar & Broth) and sterilization. I have the crystalized form of this supplement but I am trying to determine how to add the 0.05% Cysteine (0.5g/L) recommendations. So if anyone has a protocol that I can work with of guide addition of L-Cysteine, it will be very helpful to my research. I anticipate your helpful contributions.
Thank you.
Certainly! Here's a detailed protocol for preparing Bifidobacteria media (Agar and Broth) with the addition of L-cysteine HCl at a concentration of 0.05% (0.5g/L):
Bifidobacteria Agar with L-Cysteine HCl:
Ingredients:
- Bifidobacteria Agar Base
- L-Cysteine HCl (crystalline form) - 0.5g/L
- Distilled water
- Appropriate supplements (if needed)
Protocol:
Bifidobacteria Broth with L-Cysteine HCl:
Ingredients:
- Bifidobacteria Broth Base
- L-Cysteine HCl (crystalline form) - 0.5g/L
- Distilled water
- Appropriate supplements (if needed)
Protocol:
Sterilization and Storage:
- Autoclave both the agar and broth media at 121°C for 15-20 minutes for sterilization.
- Allow the media to cool to around 45-50°C before use or storage.
- Store the prepared media in a cool, dark place or refrigerate if not used immediately. Check the expiration date of the media and discard if expired.
This protocol provides a step-by-step guide for preparing Bifidobacteria media with the addition of L-Cysteine HCl at a concentration of 0.05% (0.5g/L). Adjustments can be made based on your specific experimental needs and the recommendations provided by the media manufacturer or relevant literature.
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