Exeter/2 July 2013

Learnt how to make LB broth and agar

For one litre of broth:

- 10 g tryptone

- 5 g yeast extract

- 5 g NaCl

These were weighed out using a standard tabletop scale, accurate to 2 decimal places.

They were added to ~800 ml of distilled water and mixed using a magnetic stirrer. When dissolved, more distilled water was added to bring the overall volume to 1000 ml (1 litre)

This was poured into 200 ml Duran bottles, and 3g of agar powder was added to each. This will not dissolve until the bottle is autoclaved for 20 minutes at 120°C.

We also made several 50 ml batches of LB broth, stored in conical flasks.

Learnt how to make SOB growth media

For one litre of SOB:

- 5 g yeast extract

-20 g tryptone

- 0.584 g NaCl

- 0.186 g KCl

- 2.4 g MgSO4

The tryptone, yeast extract and MgSO4 were weighed out using a standard tabletop scale, accurate to 2 decimal places. The NaCl and KCl were weighed out using a scale accurate to 4 decimal places.

This was dissolved into ~800 ml of distilled water using a magnetic stirrer, then more distilled water was added until the overall volume was 1 litre.

Made 20% weight by volume glucose solution

20g powdered glucose was added to 20 ml distilled water. The overall volume was made up to 100 ml by adding more distilled water. This was dissolved by using a magnetic stirrer.

Filter sterilised the solution, and store in two 50 ml Falcon tubes.

Take me back to the notebook.