(1 vs. 7) ProtocolsRecipesS2 < Lab

Revision 72013-09-05 - GabrielSuarez

S2

Used for Acinetobacter

Changed:

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1L	5L	Component
1.5g	7.5 g	KH₂PO₄
25.5g	127.5 g	Na₂HPO₄ 7H₂O

>
>

Recipe for 900mL

(Autoclave in three separate bottles - 300mL each)

Deleted:

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2g

10 g

NH₄Cl

Changed:

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After autoclaving, wait for containers to return to room temperature, then add the following sterile ingredients.

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Bottle 1 (Erlenmeyer flask: 300mL)

Added:

>
>

300mL	Component
1.35g	KH₂PO₄ (monobasic)
22.95g	Na₂HPO₄ 7H₂O

Changed:

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1L	Component
1ml	10% MgSO₄
0.09ul	1M CaCl₂

>
>

Bottle 2 (Erlenmeyer flask: 300mL)

300mL	Component
0.09g	MgSO₄

Deleted:

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0.5ml	0.1% FeSO₄ 7H₂O
5 ml	90% Lactic Acid

Added:

>
>

Bottle 3 (1L bottle: 300mL)

300mL	Component
1.8g	NH₄Cl
81ul	1M CaCl₂

After autoclaving, let these reach room temperature and transfer all into one 1L bottle (Bottle 3), then, finally add the following sterile components:

0.45ml	0.1% FeSO₄ 7H₂O
4.5 ml	90% Lactic Acid

The functional constraints are that Calcium Chloride cannot be autoclaved in the presence of sulfates, or it will precipitate as calcium sulfate. Ferrous sulfate when autoclaved will decompose to ferric sulfate, ferric oxide and sulfur dioxide. This means that either calcium chloride OR magnesium sulfate may be added before autoclaving, but not both.

Final composition
190.2 mM	Sodium (Na⁺)
11.0 mM	Potassium (K⁺)
37.8 mM	Ammonium (NH₄⁺)
0.09 mM	Calcium (Ca²⁺)
0.83 mM	Magnesium (Mg²⁺)
0.033 mM	Iron II (Fe²⁺)
38.0 mM	Chloride (Cl^-)
0.87 mM	Sulfate (SO₄^2-)
106.1 mM	Phosphate (PO₄^3-)
50.0 mM	Lactic Acid

Changed:

<
<

-- Main.AurkoDasgupta - 03 Jun 2013

>
>

-- Main.GabrielSuarez - 05 Sep 2013

Revision 62013-06-27 - AurkoDasgupta

S2

Used for Acinetobacter

1L	5L	Component
1.5g	7.5 g	KH₂PO₄
25.5g	127.5 g	Na₂HPO₄ 7H₂O
2g	10 g	NH₄Cl

Changed:

<
<

After autoclaving, add the following sterile ingredients.

>
>

After autoclaving, wait for containers to return to room temperature, then add the following sterile ingredients.

1L	Component
1ml	10% MgSO₄
0.09ul	1M CaCl₂
0.5ml	0.1% FeSO₄ 7H₂O
5 ml	90% Lactic Acid

The functional constraints are that Calcium Chloride cannot be autoclaved in the presence of sulfates, or it will precipitate as calcium sulfate. Ferrous sulfate when autoclaved will decompose to ferric sulfate, ferric oxide and sulfur dioxide. This means that either calcium chloride OR magnesium sulfate may be added before autoclaving, but not both.

Final composition
190.2 mM	Sodium (Na⁺)
11.0 mM	Potassium (K⁺)

Changed:

<
<

37.8 mM

Ammonium (NH₄)

>
>

37.8 mM

Ammonium (NH₄⁺)

0.09 mM	Calcium (Ca²⁺)
0.83 mM	Magnesium (Mg²⁺)

Added:

>
>

0.033 mM

Iron II (Fe²⁺)

38.0 mM	Chloride (Cl^-)
0.87 mM	Sulfate (SO₄^2-)
106.1 mM	Phosphate (PO₄^3-)
50.0 mM	Lactic Acid

-- Main.AurkoDasgupta - 03 Jun 2013

Revision 52013-06-27 - AurkoDasgupta

S2

Used for Acinetobacter

1L	5L	Component
1.5g	7.5 g	KH₂PO₄
25.5g	127.5 g	Na₂HPO₄ 7H₂O

Deleted:

<
<

0.1g

0.5 g

MgSO₄

2g

10 g

NH₄Cl

Deleted:

<
<

0.01g	0.05 g	CaCl₂
0.0005g	0.0025 g	FeSO₄ 7H₂O

Added:

>
>

After autoclaving, add the following sterile ingredients.

Changed:

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<

To each 1L, add 5ml of 85-90% lactic acid. Bring final pH to 6.65.

>
>

1L	Component

Added:

>
>

1ml	10% MgSO₄
0.09ul	1M CaCl₂
0.5ml	0.1% FeSO₄ 7H₂O
5 ml	90% Lactic Acid

Changed:

<
<

Calcium Chloride may be better added as 1ml of 1% solution, or 90ul of 1M solution.

>
>

The functional constraints are that Calcium Chloride cannot be autoclaved in the presence of sulfates, or it will precipitate as calcium sulfate. Ferrous sulfate when autoclaved will decompose to ferric sulfate, ferric oxide and sulfur dioxide. This means that either calcium chloride OR magnesium sulfate may be added before autoclaving, but not both.

Deleted:

<
<

Ferrous Sulfate may be better added as 0.5ml of a 0.1% solution.

It might be best to autoclave the calcium chloride separate from the sulfates, as there seems to be a risk of calcium sulfate precipitate otherwise.

Final composition
190.2 mM	Sodium (Na⁺)
11.0 mM	Potassium (K⁺)
37.8 mM	Ammonium (NH₄)
0.09 mM	Calcium (Ca²⁺)
0.83 mM	Magnesium (Mg²⁺)

Changed:

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<

38.0 mM

Chloride (Cl^-

>
>

38.0 mM

Chloride (Cl^-)

0.87 mM	Sulfate (SO₄^2-)
106.1 mM	Phosphate (PO₄^3-)
50.0 mM	Lactic Acid

-- Main.AurkoDasgupta - 03 Jun 2013

Revision 42013-06-25 - AurkoDasgupta

S2

Used for Acinetobacter

1L	5L	Component
1.5g	7.5 g	KH₂PO₄
25.5g	127.5 g	Na₂HPO₄ 7H₂O
0.1g	0.5 g	MgSO₄
2g	10 g	NH₄Cl
0.01g	0.05 g	CaCl₂
0.0005g	0.0025 g	FeSO₄ 7H₂O

To each 1L, add 5ml of 85-90% lactic acid. Bring final pH to 6.65.

Calcium Chloride may be better added as 1ml of 1% solution, or 90ul of 1M solution.

Ferrous Sulfate may be better added as 0.5ml of a 0.1% solution.

Added:

>
>

It might be best to autoclave the calcium chloride separate from the sulfates, as there seems to be a risk of calcium sulfate precipitate otherwise.

Final composition

Changed:

<
<

95.1 mM

Sodium (Na⁺)

>
>

190.2 mM

Sodium (Na⁺)

11.0 mM	Potassium (K⁺)
37.8 mM	Ammonium (NH₄)
0.09 mM	Calcium (Ca²⁺)
0.83 mM	Magnesium (Mg²⁺)
38.0 mM	Chloride (Cl^-
0.87 mM	Sulfate (SO₄^2-)
106.1 mM	Phosphate (PO₄^3-)
50.0 mM	Lactic Acid

-- Main.AurkoDasgupta - 03 Jun 2013

Revision 32013-06-21 - AurkoDasgupta

S2

Used for Acinetobacter

1L	5L	Component
1.5g	7.5 g	KH₂PO₄
25.5g	127.5 g	Na₂HPO₄ 7H₂O
0.1g	0.5 g	MgSO₄
2g	10 g	NH₄Cl
0.01g	0.05 g	CaCl₂
0.0005g	0.0025 g	FeSO₄ 7H₂O

To each 1L, add 5ml of 85-90% lactic acid. Bring final pH to 6.65.

Calcium Chloride may be better added as 1ml of 1% solution, or 90ul of 1M solution.

Ferrous Sulfate may be better added as 0.5ml of a 0.1% solution.

Added:

>
>

Final composition
95.1 mM	Sodium (Na⁺)
11.0 mM	Potassium (K⁺)
37.8 mM	Ammonium (NH₄)
0.09 mM	Calcium (Ca²⁺)
0.83 mM	Magnesium (Mg²⁺)
38.0 mM	Chloride (Cl^-
0.87 mM	Sulfate (SO₄^2-)
106.1 mM	Phosphate (PO₄^3-)
50.0 mM	Lactic Acid

-- Main.AurkoDasgupta - 03 Jun 2013

Revision 22013-06-04 - AurkoDasgupta

S2

Used for Acinetobacter

1L	5L	Component
1.5g	7.5 g	KH₂PO₄
25.5g	127.5 g	Na₂HPO₄ 7H₂O
0.1g	0.5 g	MgSO₄
2g	10 g	NH₄Cl
0.01g	0.05 g	CaCl₂

Added:

>
>

0.0005g

0.0025 g

FeSO₄ 7H₂O

Deleted:

<
<

To each 1L, each 5ml of 85-90% lactic acid. Bring final pH to 6.65.

Changed:

<
<

Calcium Chloride may be better added as 1ml of 1% solution.

>
>

To each 1L, add 5ml of 85-90% lactic acid. Bring final pH to 6.65.

Added:

>
>

Calcium Chloride may be better added as 1ml of 1% solution, or 90ul of 1M solution.

Ferrous Sulfate may be better added as 0.5ml of a 0.1% solution.

-- Main.AurkoDasgupta - 03 Jun 2013

Revision 12013-06-03 - AurkoDasgupta

S2

Used for Acinetobacter

1L	5L	Component
1.5g	7.5 g	KH₂PO₄
25.5g	127.5 g	Na₂HPO₄ 7H₂O
0.1g	0.5 g	MgSO₄
2g	10 g	NH₄Cl
0.01g	0.05 g	CaCl₂

To each 1L, each 5ml of 85-90% lactic acid. Bring final pH to 6.65.

Calcium Chloride may be better added as 1ml of 1% solution.

-- Main.AurkoDasgupta - 03 Jun 2013

Difference: ProtocolsRecipesS2 (1 vs. 7)

Revision 72013-09-05 - GabrielSuarez

S2

Revision 62013-06-27 - AurkoDasgupta

S2

Revision 52013-06-27 - AurkoDasgupta

S2

Revision 42013-06-25 - AurkoDasgupta

S2

Revision 32013-06-21 - AurkoDasgupta

S2

Revision 22013-06-04 - AurkoDasgupta

S2

Revision 12013-06-03 - AurkoDasgupta

S2