Oct 18, 2024
Luria Bertani Broth and agarized
- Marta Puddu1
- 1Polimi
Protocol Citation: Marta Puddu 2024. Luria Bertani Broth and agarized. protocols.io https://dx.doi.org/10.17504/protocols.io.j8nlk87o6l5r/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: October 17, 2024
Last Modified: October 18, 2024
Protocol Integer ID: 110173
Abstract
Recipe and protocol for Luria Bertani broth (Miller version) preparation, an optional agarization step is provided.
Recipe and steps to prepare your mixture
Recipe and steps to prepare your mixture
Take note of the position of YourFavouriteIngredients and take them from the reagent-room.
5m
Weight ingredients:
Tryptone powder (technical grade) 10 g/L
Yeast extract 5 g/L
NaCl (Miller recipe) 10 g/L
Milli Q or Tap water to volume 1L.
10m
Add all the solid ingredients to a plastic or glass becker, along with 600-700mL of water to dissolve the ingredients.
Its facultative to measure Water with a cilinder.
Its facultative to add autoclavable components (vitamins, antibiotics etc.)
5m
Agarization
Agarization
2m
2m
Add 15 g/L Agar microbiology grade along with 100mL of water to "dissolve" the agar. It will settle to the bottom due to low solubilization below 50°C, it will be mixed during autoclaving step.
Finalise your broth
Finalise your broth
1m
1m
Make up to final 1L volume with remaining water and further mix.
1m
Autoclave it!
Autoclave it!
15m
15m
Pour the content into a Duran to autoclave the mixture.
Do not fill the whole bottle, leave some headspace (1/4 of total volume HS).
Do not seal the bottle, leave the top half-round open.
Remember to place autoclave-tape over your bottles.
Autoclave setting: 121°C for 15min, 15psi/1bar
5m