Team:Yale/Our Project/Protocols/minimal media

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Revision as of 15:57, 24 October 2010

iGEM Yale

protocols

Morpholinepropanesulfonic Acid (MOPS) Minimal Medium Recipe

To make 1 liter of the 10x concentrate combine the following in order
1. 400 mL of 1.0 M morpholinopropane sulfonate (MOPS) adjusted to pH 7.4 with KOH
2. 40 mL of 1.0 M Tricine adjusted to pH 7.4 with KOH
3. 10 mL of .01 M FeSO4
4. 50 mL of 1.90 M NH4Cl
5. 10 mL of 0.276 M K2SO4
6. 10 mL of 5.0 * 10-4 M CaCl2 (Start at 100x and then dilute)
7. 10 mL of 0.528 M MgCl2
8. 100 mL of 5.0 M NaCl
9. 10 mL of micronutrient solution made by putting the following in 100 mL and then making two hundredfold dilutions in series
  • a. 3 * 10-3 moles of (NH4)6(Mo7)24
  • b. 0.4 moles of H3BO3
  • c. 0.03 moles of CoCl2
  • d. 0.01 moles of CuSO4
  • e. 0.08 moles of MnCl2
  • f. 0.01 moles of ZnSO4
10. 360 mL of glass-distilled water
Filter sterilize the concentrate with a filter pre-rinsed in the solution –the result can be stored for long periods of time To create medium at normal concentration combine the following—result should be have pH near 7.2 1. 100 mL MOPS concentrate
2. 10 mL of 0.132 M glycerol 2-phosphate
3. 10 mL of carbon source
4. 880 mL of glass-distilled water, autoclaved

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