(r2) Barrick Lab :: ProtocolsRecipesABMS

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---+++ ABMS: Acinetobacter Minimal Succinate

Courtesy of the Averhoff lab, with modifications for available reagents.

To make standard ABMS, combine the following sterilized ingredients at room temperature:
|  *1L*  |  *5L*  |  *Component*  |
| 880mL | 4.4L | diH<sub>2</sub>O |
| 20mL | 100mL | 1M Sodium Succinate |
| 50mL | 250mL | 20X Mineral Solution |
| 50mL | 250mL | 20X Phosphate Buffer |

To make ABMS plates:

Add 16g/L agar and 1mL/L antifoam to diH<sub>2</sub>O, then autoclave in a 2L flask. When water and agar are cool enough to touch, add component solutions. Optional: warm components at 55C in a water bath before combining with agar to reduce the temperature difference when combined.

---++++ ABMS Component Recipes

*SL9 (trace minerals)*
|  *1L*  |  *Component*  |
| 800mL | diH<sub>2</sub>O |
| 12.8g | Nitrotriacetic acid |
| 2g | FeSO<sub>4</sub>•7H<sub>2</sub>O (note: first dissolve in 2M HCl) |
| 104mg | CoCl<sub>2</sub> (anhydrous) |
| 122mg | MnCl<sub>2</sub>•4H<sub>2</sub>O |
| 70mg | ZnCl<sub>2</sub> |
| 36mg | NaMoO<sub>4</sub>•2H<sub>2</sub>O |
| 13mg | NiCl<sub>2</sub> |

Add 1mL of 10mL 10X solution:
| 60mg | H<sub>3</sub>BO<sub>3</sub> |
| 20mg | CuCl<sub>2</sub>•2H<sub>2</sub>O | 

Bring to 1000mL and pH 6.5. Note: this requires a lot of NaOH, so leave room when bringing up to volume.

*Mineral Solution (20X)*
| *500mL*  |  *Component*  |
| 10g | NH<sub>4</sub>Cl |
| 5.9g | MgSO<sub>4</sub>•7H<sub>2</sub>O |
| 1g | KNO<sub>3</sub> | 
| 20mg | (NH<sub>4</sub>)<sub>6</sub>Mo<sub>7</sub>O<sub>24</sub>•4H<sub>2</sub>O |
| 931.5µL | CaCl<sub>2</sub> Solution |
| 10mL | SL9 |

Bring to 500mL in a graduated cylinder with deionized water.

*Phosphate Buffer (20X)*
|  *500mL*  |  *Components*  |
| 68g | KH<sub>2</sub>PO<sub>4</sub> |
| 132.5g | Na<sub>2</sub>HPO<sub>4</sub>•H<sub>2</sub>O |
| 400mL | Deionized water, pH 6.8 |

Bring to 500mL.

Barrick Lab > ProtocolsMediaRecipes > ProtocolsRecipesABMS

Contributors to this topic

JeffreyBarrick, GabrielSuarez, KyleDugan

Topic revision: r2 - 2016-07-20 - 19:49:09 - Main.KyleDugan