<noautolink> ---+++ ABMS: Acinetobacter Minimal Succinate Courtesy of the Averhoff lab, with modifications for available reagents. To make standard ABMS, combine the following *pre-sterilized* ingredients at room temperature: | *1L* | *5L* | *Component* | | 880 mL | 4.4 L | diH<sub>2</sub>O | | 20 mL | 100 mL | 1M Sodium Succinate | | 50 mL | 250 mL | 20× Mineral Solution | | 50 mL | 250 mL | 20× Phosphate Buffer | To make ABMS agar: Per liter of media, add 16 g/L agar and 1 mL/L antifoam to diH<sub>2</sub>O, then autoclave in a 2 L flask. When water and agar are cool enough to touch, add component solutions. Optional: warm components at 55°C in a water bath before combining with agar to reduce the temperature difference when combined. ---++++ ABMS Component Recipes *SL9 (trace minerals)* | *1L* | *Component* | | 800 mL | diH<sub>2</sub>O | | 12.8 g | Nitrilotriacetic acid | | 2 g | FeSO<sub>4</sub>•7H<sub>2</sub>O (note: first dissolve in 2M HCl) | | 104 mg | CoCl<sub>2</sub> (anhydrous) | | 122 mg | MnCl<sub>2</sub>•4H<sub>2</sub>O | | 70 mg | ZnCl<sub>2</sub> | | 36 mg | Na<sub>2</sub>MoO<sub>4</sub>•2H<sub>2</sub>O | | 13 mg | NiCl<sub>2</sub> | Add 1 mL of 10 mL 10× solution: | 60 mg | H<sub>3</sub>BO<sub>3</sub> | | 20 mg | CuCl<sub>2</sub>•2H<sub>2</sub>O | Adjust pH to 6.5 using 1 M NaOH. Bring volume to 1000 mL with diH<sub>2</sub>O. Note: this requires a lot of NaOH, so leave room when bringing up to volume. *Mineral Solution (20×)* | *500 mL* | *Component* | | 10 g | NH<sub>4</sub>Cl | | 5.8 g | MgSO<sub>4</sub>•7H<sub>2</sub>O | | 1 g | KNO<sub>3</sub> | | 0.67 g | CaCl<sub>2</sub>•2H<sub>2</sub>O † | | 20 mg | (NH<sub>4</sub>)<sub>6</sub>Mo<sub>7</sub>O<sub>24</sub>•4H<sub>2</sub>O | | 10 mL | SL 9 | *† Substitution:* You may use 465.8 µL of a 1 M CaCl<sub>2</sub> per 500 ml of stock solution for this component. The final concentration of Ca<sup>2+</sup> in the 20× stock is 9.12 mM. Bring to 500mL in a graduated cylinder with deionized water. *Phosphate Buffer (20×)* | *500mL* | *Components* | | 68 g | KH<sub>2</sub>PO<sub>4</sub> | | 132.5 g | Na<sub>2</sub>HPO<sub>4</sub>•H<sub>2</sub>O | Bring to 500 mL with diH<sub>2</sub>O; pH should be ~6.9-7.0 (you may add 400 mL diH<sub>2</sub>O first and bring pH up to 6.8 then complete volume to 500mL) * Source: Averhoff lab
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Topic revision: r6 - 2019-06-23 - JeffreyBarrick
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