Exeter/2 July 2013

From 2013.igem.org

Learn how to make LB broth and agar

For one litre of broth we need...

- 10g tryptone

- 5g yeast extract

- 5g NaCl

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

Add to ~800ml of distilled water and mix using a magnetic stirrer. When dissolved, add more distilled water to bring the overall amount to 1000ml (1 litre)

Pour into 200ml Duran bottles, and add 3g of agar powder to each. This will not dissolve until the bottle is autoclaved for 20 minutes at 120oC.

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

Learn how to make SOB growth media

For one litre of SOB we need...

- 5g yeast extract

-20g tryptone

- 0.584g NaCl

- 0.186g KCl

- 2.4g 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.

Dissolve into ~800ml of distilled water using a magnetic stirrer, then add more distilled water until the overall volume is 1 litre.

Make 20% weight by volume glucose solution

Add 20g powdered glucose to 20ml distilled water. Make up overall volume to 100ml by adding more distilled water. Dissolve by using magnetic stirrer.

Filter sterilise the solution and store in 2 50ml Falcon tube.