Difference: ProtocolsRecipesABMS (1 vs. 6)

Revision 62019-06-23 - JeffreyBarrick

 
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
Changed:
<
<
50 mL 250 mL 20X Mineral Solution
50 mL 250 mL 20X Phosphate Buffer
>
>
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 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
Changed:
<
<
12.8 g Nitrotriacetic acid
>
>
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
Changed:
<
<
36 mg NaMoO4•2H2O
>
>
36 mg Na2MoO4•2H2O
 
13 mg NiCl2
Changed:
<
<		Add 1 mL of 10 mL 10X solution:
>
>		Add 1 mL of 10 mL 10× solution:
 
60 mg H3BO3
20 mg CuCl2•2H2O
Changed:
<
<		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.
 
Changed:
<
<		Mineral Solution (20X)
>
>		Mineral Solution (20×)
 
500 mL Component
10 g NH4Cl
Changed:
<
<
5.9 g MgSO4•7H2O
>
>
5.8 g MgSO4•7H2O
 
1 g KNO3
Added:
>
>
0.67 g CaCl2•2H2O †
 
20 mg (NH4)6Mo7O24•4H2O
Changed:
<
<
931.5 µL CaCl2 Solution (1M) - SigmaAldrich
>
>
10 mL SL 9
Deleted:
<
<
10 mL SL9
 
Added:
>
>		† 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.
Changed:
<
<		Phosphate Buffer (20X)
>
>		Phosphate Buffer (20×)
 
500mL Components
68 g KH2PO4
132.5 g Na2HPO4•H2O
Changed:
<
<		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)
Added:
>
>		 * Source: Averhoff lab
 

Revision 52017-11-09 - GabrielSuarez

 
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
50 mL 250 mL 20X Mineral Solution
50 mL 250 mL 20X 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 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 Nitrotriacetic acid
2 g FeSO4•7H2O (note: first dissolve in 2M HCl)
104 mg CoCl2 (anhydrous)
122 mg MnCl2•4H2O
70 mg ZnCl2
36 mg NaMoO4•2H2O
13 mg NiCl2

Add 1 mL of 10 mL 10X solution:

60 mg H3BO3
20 mg CuCl2•2H2O

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.

Mineral Solution (20X)

500 mL Component
10 g NH4Cl
5.9 g MgSO4•7H2O
1 g KNO3
20 mg (NH4)6Mo7O24•4H2O
931.5 µL CaCl2 Solution (1M) - SigmaAldrich
10 mL SL9

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

Phosphate Buffer (20X)

500mL Components
68 g KH2PO4
132.5 g Na2HPO4•H2O
Changed:
<
<		Bring to 500 mL with diH2O.
>
>		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)
 

Revision 42017-11-09 - GabrielSuarez

 
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
50 mL 250 mL 20X Mineral Solution
50 mL 250 mL 20X 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 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 Nitrotriacetic acid
Changed:
<
<
2 g FeSO4•7H2O (note: first dissolve in 2M HCl)
>
>
2 g FeSO4•7H2O (note: first dissolve in 2M HCl)
 
104 mg CoCl2 (anhydrous)
Changed:
<
<
122 mg MnCl2•4H2O
>
>
122 mg MnCl2•4H2O
 
70 mg ZnCl2
Changed:
<
<
36 mg NaMoO4•2H2O
>
>
36 mg NaMoO4•2H2O
 
13 mg NiCl2

Add 1 mL of 10 mL 10X solution:

60 mg H3BO3
Changed:
<
<
20 mg CuCl2•2H2O
>
>
20 mg CuCl2•2H2O
  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.

Mineral Solution (20X)

500 mL Component
10 g NH4Cl
Changed:
<
<
5.9 g MgSO4•7H2O
1 g KNO3
20 mg (NH4)6Mo7O24•4H2O
931.5 µL CaCl2 Solution
>
>
5.9 g MgSO4•7H2O
1 g KNO3
20 mg (NH4)6Mo7O24•4H2O
931.5 µL CaCl2 Solution (1M) - SigmaAldrich
 
10 mL SL9

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

Phosphate Buffer (20X)

500mL Components
68 g KH2PO4
Changed:
<
<
132.5 g Na2HPO4•H2O
>
>
132.5 g Na2HPO4•H2O
  Bring to 500 mL with diH2O.

Revision 32016-07-21 - JeffreyBarrick

 
META TOPICPARENT name="ProtocolsMediaRecipes"

ABMS: Acinetobacter Minimal Succinate

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

Changed:
<
<		To make standard ABMS, combine the following sterilized ingredients at room temperature:
>
>		To make standard ABMS, combine the following pre-sterilized ingredients at room temperature:
 
1L 5L Component
Changed:
<
<
880mL 4.4L diH2O
20mL 100mL 1M Sodium Succinate
50mL 250mL 20X Mineral Solution
50mL 250mL 20X Phosphate Buffer
>
>
880 mL 4.4 L diH2O
20 mL 100 mL 1M Sodium Succinate
50 mL 250 mL 20X Mineral Solution
50 mL 250 mL 20X Phosphate Buffer
 
Changed:
<
<		To make ABMS plates:
>
>		To make ABMS agar:
 
Changed:
<
<		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.
>
>		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
Changed:
<
<
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
>
>
800 mL diH2O
12.8 g Nitrotriacetic acid
2 g FeSO4•7H2O (note: first dissolve in 2M HCl)
104 mg CoCl2 (anhydrous)
122 mg MnCl2•4H2O
70 mg ZnCl2
36 mg NaMoO4•2H2O
13 mg NiCl2
 
Changed:
<
<		Add 1mL of 10mL 10X solution:
60mg H3BO3
20mg CuCl2•2H2O
>
>		Add 1 mL of 10 mL 10X solution:
60 mg H3BO3
20 mg CuCl2•2H2O
 
Changed:
<
<		Bring to 1000mL and pH 6.5. 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 1000mL with diH2O. Note: this requires a lot of NaOH, so leave room when bringing up to volume.
  Mineral Solution (20X)
Changed:
<
<
500mL Component
10g NH4Cl
5.9g MgSO4•7H2O
1g KNO3
20mg (NH4)6Mo7O24•4H2O
931.5µL CaCl2 Solution
10mL SL9
>
>
500 mL Component
10 g NH4Cl
5.9 g MgSO4•7H2O
1 g KNO3
20 mg (NH4)6Mo7O24•4H2O
931.5 µL CaCl2 Solution
10 mL SL9
  Bring to 500mL in a graduated cylinder with deionized water.

Phosphate Buffer (20X)

500mL Components
Changed:
<
<
68g KH2PO4
132.5g Na2HPO4•H2O
>
>
68 g KH2PO4
132.5 g Na2HPO4•H2O
Deleted:
<
<
400mL Deionized water, pH 6.8
 
Changed:
<
<		Bring to 500mL.
>
>		Bring to 500 mL with diH2O.
 

Revision 22016-07-20 - KyleDugan

 
META TOPICPARENT name="ProtocolsMediaRecipes"
Changed:
<
<

ABMS

>
>

ABMS: Acinetobacter Minimal Succinate

 
Changed:
<
<		ABMS stands for.... It is used for....
>
>		Courtesy of the Averhoff lab, with modifications for available reagents.
 
Changed:
<
<
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
>
>		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
Deleted:
<
<
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.
 
Changed:
<
<		If making ABMS solid media also add 15 g of agar and 1 ml of 5% antifoam per liter.
>
>		To make ABMS plates:
Added:
>
>		 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.

 

Revision 12016-07-20 - JeffreyBarrick

 
META TOPICPARENT name="ProtocolsMediaRecipes"

ABMS

ABMS stands for.... It is used for....

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
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.

If making ABMS solid media also add 15 g of agar and 1 ml of 5% antifoam per liter.

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