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| META TOPICPARENT |
name="ProtocolsMediaRecipes" |
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< | ABMS |
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> | ABMS: Acinetobacter Minimal Succinate |
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<
< | ABMS stands for.... It is used for.... |
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> | Courtesy of the Averhoff lab, with modifications for available reagents. |
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< |
| 1L |
5L |
Component |
| 0.5 g |
2.5 g |
Yeast Extract |
| 0.5 g |
2.5 g |
Proteose Peptone No. 3 |
| 0.5 g |
2.5 g |
Casamino Acids |
| 0.5 g |
2.5 g |
Dextrose |
| 0.5 g |
2.5 g |
Soluble Starch |
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>
> | To make standard ABMS, combine the following sterilized ingredients at room temperature:
| 1L |
5L |
Component |
| 880mL |
4.4L |
diH2O |
| 20mL |
100mL |
1M Sodium Succinate |
| 50mL |
250mL |
20X Mineral Solution |
| 50mL |
250mL |
20X Phosphate Buffer |
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<
< |
| 0.3 g |
1.5 g |
Sodium Pyruvate |
| 0.3 g |
1.5 g |
Dipotassium Phosphate |
| 0.05 g |
0.25 g |
Magnesium Sulfate |
Add dH2O to final volume and autoclave. |
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< | If making ABMS solid media also add 15 g of agar and 1 ml of 5% antifoam per liter. |
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> | To make ABMS plates: |
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>
> |
Add 16g/L agar and 1mL/L antifoam to diH2O, then autoclave in a 2L 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 |
| 800mL |
diH2O |
| 12.8g |
Nitrotriacetic acid |
| 2g |
FeSO4•7H2O (note: first dissolve in 2M HCl) |
| 104mg |
CoCl2 (anhydrous) |
| 122mg |
MnCl2•4H2O |
| 70mg |
ZnCl2 |
| 36mg |
NaMoO4•2H2O |
| 13mg |
NiCl2 |
Add 1mL of 10mL 10X solution:
| 60mg |
H3BO3 |
| 20mg |
CuCl2•2H2O |
Bring to 1000mL and pH 6.5. Note: this requires a lot of NaOH, so leave room when bringing up to volume.
Mineral Solution (20X)
| 500mL |
Component |
| 10g |
NH4Cl |
| 5.9g |
MgSO4•7H2O |
| 1g |
KNO3 |
| 20mg |
(NH4)6Mo7O24•4H2O |
| 931.5µL |
CaCl2 Solution |
| 10mL |
SL9 |
Bring to 500mL in a graduated cylinder with deionized water.
Phosphate Buffer (20X)
| 500mL |
Components |
| 68g |
KH2PO4 |
| 132.5g |
Na2HPO4•H2O |
| 400mL |
Deionized water, pH 6.8 |
Bring to 500mL. |
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