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 diH
2O, 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 |
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 diH
2O. 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 µL of a 1 M CaCl
2 per 500 ml of stock solution for this component. The final concentration of Ca
2+ 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 diH
2O; pH should be ~6.9-7.0 (you may add 400 mL diH
2O first and bring pH up to 6.8 then complete volume to 500mL)
* Source: Averhoff lab
Contributors to this topic
JeffreyBarrick, GabrielSuarez, KyleDugan
Topic revision: r6 - 2019-06-23 - 12:09:45 - Main.JeffreyBarrick