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META TOPICPARENT |
name="ProtocolsMediaRecipes" |
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 |
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|
< < |
50 mL |
250 mL |
20X Mineral Solution |
50 mL |
250 mL |
20X Phosphate Buffer |
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> > |
50 mL |
250 mL |
20× Mineral Solution |
50 mL |
250 mL |
20× Phosphate Buffer |
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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 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 |
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|
< < |
12.8 g |
Nitrotriacetic acid |
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> > |
12.8 g |
Nitrilotriacetic acid |
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|
2 g |
FeSO4•7H2O (note: first dissolve in 2M HCl) |
104 mg |
CoCl2 (anhydrous) |
122 mg |
MnCl2•4H2O |
70 mg |
ZnCl2 |
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< < |
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> > |
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< < | Add 1 mL of 10 mL 10X solution: |
> > | Add 1 mL of 10 mL 10× solution: |
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60 mg |
H3BO3 |
20 mg |
CuCl2•2H2O |
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< < | 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. |
> > | 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. |
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< < | Mineral Solution (20X) |
> > | Mineral Solution (20×) |
|
500 mL |
Component |
10 g |
NH4Cl |
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|
< < |
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> > |
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> > |
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|
< < |
931.5 µL |
CaCl2 Solution (1M) - SigmaAldrich |
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> > |
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< < |
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> > | † Substitution: You may use 465.8 µL 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. |
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< < | Phosphate Buffer (20X) |
> > | Phosphate Buffer (20×) |
|
500mL |
Components |
68 g |
KH2PO4 |
132.5 g |
Na2HPO4•H2O |
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< < | 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) |
> > | 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) |
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> > |
* Source: Averhoff lab |
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