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

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 Stir to mix
 * 1) Weigh out 2.7g Ferric Chloride
 * 2) Weigh out 2.1g Citric Acid
 * 3) Add 10ml distilled water



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





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




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