Difference: ProtocolsRecipesS2 (1 vs. 7)

Revision 72013-09-05 - GabrielSuarez

 

S2

Used for Acinetobacter

Changed:
<
<
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:
<
<
2g 10 g NH4Cl
 
Changed:
<
<		After autoclaving, wait for containers to return to room temperature, then add the following sterile ingredients.
>
>		Bottle 1 (Erlenmeyer flask: 300mL)
Added:
>
>
300mL Component
1.35g KH2PO4 (monobasic)
22.95g Na2HPO4 7H2O
 
Changed:
<
<
1L Component
1ml 10% MgSO4
0.09ul 1M CaCl2
>
>		Bottle 2 (Erlenmeyer flask: 300mL)
300mL Component
0.09g MgSO4
Deleted:
<
<
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 (SO42-)
106.1 mM Phosphate (PO43-)
50.0 mM Lactic Acid
Changed:
<
<		-- AurkoDasgupta - 03 Jun 2013
>
>		-- 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 (SO42-)
106.1 mM Phosphate (PO43-)
50.0 mM Lactic Acid

-- 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:
<
<		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:
<
<
38.0 mM Chloride (Cl-
>
>
38.0 mM Chloride (Cl-)
 
0.87 mM Sulfate (SO42-)
106.1 mM Phosphate (PO43-)
50.0 mM Lactic Acid

-- 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 (SO42-)
106.1 mM Phosphate (PO43-)
50.0 mM Lactic Acid

-- 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 (SO42-)
106.1 mM Phosphate (PO43-)
50.0 mM Lactic Acid
  -- 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.

 

-- 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

View topic | History: r7 < r6 < r5 < r4 | More topic actions...
 
This site is powered by the TWiki collaboration platform Powered by PerlCopyright © 2008-2025 by the contributing authors. All material on this collaboration platform is the property of the contributing authors.
Ideas, requests, problems regarding TWiki? Send feedback