"
Team:Yale/Our Project/Protocols/minimal medium
From 2010.igem.org
(Difference between revisions)
m |
|||
| (One intermediate revision not shown) | |||
| Line 31: | Line 31: | ||
To make 1 liter of the 10x concentrate combine the following in order <br/> | To make 1 liter of the 10x concentrate combine the following in order <br/> | ||
1. 400 mL of 1.0 M morpholinopropane sulfonate (MOPS) adjusted to pH 7.4 with KOH <br/> | 1. 400 mL of 1.0 M morpholinopropane sulfonate (MOPS) adjusted to pH 7.4 with KOH <br/> | ||
| - | 2. 40 mL of 1.0 M Tricine adjusted to pH 7.4 with KOH | + | 2. 40 mL of 1.0 M Tricine adjusted to pH 7.4 with KOH <br/> |
| - | 3. 10 mL of .01 M | + | 3. 10 mL of .01 M FeSO<sub>4</sub> <br/> |
| - | 4. 50 mL of 1.90 M | + | 4. 50 mL of 1.90 M NH<sub>4</sub>Cl <br/> |
| - | 5. 10 mL of 0.276 M | + | 5. 10 mL of 0.276 M K<sub>2</sub>SO<sub>4</sub> <br/> |
| - | 6. 10 mL of 5.0 * 10-4 M | + | 6. 10 mL of 5.0 * 10<sup>-4</sup> M CaCl<sub>2</sub> (Make at 100x and then dilute)<br/> |
| - | 7. 10 mL of 0.528 M | + | 7. 10 mL of 0.528 M MgCl<sub>2</sub> <br/> |
8. 100 mL of 5.0 M NaCl | 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 <br/> | 9. 10 mL of micronutrient solution made by putting the following in 100 mL and then making two hundredfold dilutions in series <br/> | ||
<ul> | <ul> | ||
| - | <li>a. 3 * 10-3 moles of ( | + | <li>a. 3 * 10<sup>-3</sup> moles of (NH<sub>4</sub>)<sub>6</sub>(Mo<sub>7</sub>)<sub>24</sub> </li> |
| - | <li>b. 0.4 moles of | + | <li>b. 0.4 moles of H<sub>3</sub>BO<sub>3</sub></li> |
| - | <li>c. 0.03 moles of | + | <li>c. 0.03 moles of CoCl<sub>2</sub></li> |
| - | <li>d. 0.01 moles of | + | <li>d. 0.01 moles of CuSO>sub>4</sub> </li> |
<li>e. 0.08 moles of MnCl<sub>2</sub></li> | <li>e. 0.08 moles of MnCl<sub>2</sub></li> | ||
| - | <li>f. 0.01 moles of ZnSO<sub>4</sub> | + | <li>f. 0.01 moles of ZnSO<sub>4</sub></li> |
</ul> | </ul> | ||
10. 360 mL of glass-distilled water <br/> | 10. 360 mL of glass-distilled water <br/> | ||
Latest revision as of 19:22, 25 October 2010
our project
protocols
Morpholinepropanesulfonic acid (MOPS) minimal medium
To make 1 liter of the 10x concentrate combine the following in order1. 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 (Make 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 CuSO>sub>4
- e. 0.08 moles of MnCl2
- f. 0.01 moles of ZnSO4
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
