User:Howard Boland/Notebook/Art from Synthetic Biology/2011/01/27

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Nanomagnetic Media

I am exploring suitable media for growing nanomagnetic bacteria. I will need to make both liquid broth and agar for plates. For the broth I used ready-made Brucella broth and I also prepared Flask Standard Media. I used Kligler Iron Agar for the plates. I also tried to streak out the bacteria on standard agar plates.

Bruscella Broth

28g/L, for 200ml this yields 28g/(1000ml/200ml) = 28g/5 = 5.2g

  1. Weigh out 5.2g Bruscella Broth Powder
  2. Add 200ml distilled water

Bruscella-broth-prepared.jpg

Ferric Citrate

For the flask standard media, the last step requires ferric citrate. We did not have this in any of the labs so I tried to make some using ferric chloride and citric acid. A patent states that this is produced using 1:1 mole. Ferric Chloride (hexhydrate), a yellow toffee-like substance weighs 270.3g/mole whilst Citric Acid a sugary powder weighs 210.14g/mole.

To make 10ml 1M ferric citrate

  1. Weigh out 2.7g Ferric Chloride
  2. Weigh out 2.1g Citric Acid
  3. Add 10ml distilled water

Stir to mix

Ferric-citrate-prepared.jpg

Flask Standard Media (FSM)

The protocol for FSM was suggested in the paper (Growth and magnetosome formation by microaerophilic Magnetospirillum strains in an oxygen-controlled fermentor) for growing nanomagnetic bacteria. To make 1L FSM Media

  1. 0.1g KH2PO4 (potassium dihydrogen phosphate)
  2. 0.15g MgSO4x7H2O (magnesium sulphate)
  3. 2.38g Hepes
  4. 0.34g NaNO3 (Natrium nitrate)
  5. 0.1g Yeast Extract
  6. 3g Peptone
  7. 1ml EDTA
  8. 100µl 1M Ferric Acid
  9. Add distilled water to make up 1L

Adjust PH to 7.0

I distributed the medium in 4 bottles with 200ml media each and added the remaining to small flask using 10ml media in each.

Autoclave at 121ºC for 15 minutes

Fsm-200ml-prepared.jpg

Fsm-10ml-prepared.jpg

Kligler Iron Agar

I am experimentally using this agar to see if it produces any growth, the bottle suggests using 500g powder per 9L. To make 200ml agar, 500g/9L = 500g/9000ml => 500g/ (9000ml/200ml) = 500g/45 = 11.1g

  1. Weigh out 11.1g Powder
  2. Add 200ml distilled water

Iron-agar-prepared.jpg