Exeter/2 July 2013

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Revision as of 10:30, 16 July 2013 by JoshMiddleton (Talk | contribs)

Learn how to make LB broth and agar

For one litre of broth we need...

- 10 g tryptone

- 5 g yeast extract

- 5 g NaCl

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

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

Pour into 200 ml 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 50 ml batches of LB broth, stored in conical flasks.

Learn how to make SOB growth media

For one litre of SOB we need...

- 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.

Dissolve into ~800 ml 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 20 ml distilled water. Make up overall volume to 100 ml by adding more distilled water. Dissolve by using magnetic stirrer.

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