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 diH2O
20 mL 100 mL 1M Sodium Succinate
50 mL 250 mL 20X Mineral Solution
50 mL 250 mL 20X Phosphate Buffer

To make ABMS agar:

Per liter of media, add 16 g/L agar and 1 mL/L antifoam to diH2O, then autoclave in a 2 L 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
800 mL diH2O
12.8 g Nitrotriacetic acid
2 g FeSO4•7H2O (note: first dissolve in 2M HCl)
104 mg CoCl2 (anhydrous)
122 mg MnCl2•4H2O
70 mg ZnCl2
36 mg NaMoO4•2H2O
13 mg NiCl2

Add 1 mL of 10 mL 10X solution:

60 mg H3BO3
20 mg CuCl2•2H2O

Adjust pH to 6.5 using 1 M NaOH. Bring volume to 1000mL with diH2O. Note: this requires a lot of NaOH, so leave room when bringing up to volume.

Mineral Solution (20X)

500 mL Component
10 g NH4Cl
5.9 g MgSO4•7H2O
1 g KNO3
20 mg (NH4)6Mo7O24•4H2O
931.5 L CaCl2 Solution (1M) - SigmaAldrich
10 mL SL9

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

Phosphate Buffer (20X)

500mL Components
68 g KH2PO4
132.5 g Na2HPO4•H2O

Bring to 500 mL with diH2O; pH should be ~6.9-7.0 (you may add 400mL diH2O first and bring pH up to 6.8 then complete volume to 500mL)

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Topic revision: r5 - 09 Nov 2017 - 22:39:16 - Main.GabrielSuarez
 
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