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 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 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 Nitrilotriacetic acid
2 g FeSO4•7H2O (note: first dissolve in 2M HCl)
104 mg CoCl2 (anhydrous)
122 mg MnCl2•4H2O
70 mg ZnCl2
36 mg Na2MoO4•2H2O
13 mg NiCl2

Add 1 mL of 10 mL 10× solution:

60 mg H3BO3
20 mg CuCl2•2H2O

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

Mineral Solution (20×)

500 mL Component
10 g NH4Cl
5.8 g MgSO4•7H2O
1 g KNO3
0.67 g CaCl2•2H2O †
20 mg (NH4)6Mo7O24•4H2O
10 mL SL 9

† Substitution: You may use 465.8 L of a 1 M CaCl2 per 500 ml of stock solution for this component. The final concentration of Ca2+ 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 KH2PO4
132.5 g Na2HPO4•H2O

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

* Source: Averhoff lab

Edit | Attach | Watch | Print version | History: r6 < r5 < r4 < r3 < r2 | Backlinks | Raw View | More topic actions

 Barrick Lab  >  ProtocolsMediaRecipes  >  ProtocolsRecipesABMS

Contributors to this topic Edit topic JeffreyBarrick, GabrielSuarez, KyleDugan
Topic revision: r6 - 2019-06-23 - 12:09:45 - Main.JeffreyBarrick
This site is powered by the TWiki collaboration platform Powered by Perl This site is powered by the TWiki collaboration platformCopyright ©2023 Barrick Lab contributing authors. Ideas, requests, problems? Send feedback