ABMS: Acinetobacter Minimal Succinate

Courtesy of the Averhoff lab, with modifications for available reagents.

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

To make ABMS plates:

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 FeSO47H2O (note: first dissolve in 2M HCl)
104mg CoCl2 (anhydrous)
122mg MnCl24H2O
70mg ZnCl2
36mg NaMoO42H2O
13mg NiCl2

Add 1mL of 10mL 10X solution:

60mg H3BO3
20mg CuCl22H2O

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 MgSO47H2O
1g KNO3
20mg (NH4)6Mo7O244H2O
931.5L CaCl2 Solution
10mL SL9

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

Phosphate Buffer (20X)

500mL Components
68g KH2PO4
132.5g Na2HPO4H2O
400mL Deionized water, pH 6.8

Bring to 500mL.

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Contributors to this topic Edit topic JeffreyBarrick, GabrielSuarez, KyleDugan
Topic revision: r2 - 2016-07-20 - 19:49:09 - Main.KyleDugan
 
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