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16
A.4.1.3 KPSS
1 x KPSS (1000ml):
Solution 1
Chemical
MW (g/mol)
Conc. (mmol/l) Conc. (g/l)
KCl
74.56
123.70
9.223
MgSO
4
-
7
H
2
O
246.48
1.17
0.289
KH
2
PO
4
136.09
1.18
0.161
Solution 2
Chemical
MW (g/mol)
Conc. (mmol/l) Conc. (g/l)
CaCl
2
-
2
H
2
O
147.02
2.50
0.368
Solution 3
Chemical
MW (g/mol)
Conc. (mmol/l) Conc. (g/l)
NaHCO
3
84.01
25.00
2.100
EDTA
372.24
0.03
0.010
Glucose
198.77
5.50
1.091
1. Dissolve the chemicals in approximately 100 ml double distilled H
2
O as three individual solutions as
described in the table above. Gently heat solution 3 to dissolve the EDTA.
2. Solution 1 is added to a graduated bottle and the bottle is filled with double distilled H
2
O to a final
volume of 500 ml.
3. Solution 3 is added to the graduated bottle that is filled with additional double distilled H
2
O to a final
volume of about 850 ml.
4. Aerate the solution with carbogen (95% O
2
+ 5% CO
2
) for about 20 minutes.
5. Solution 2 is added and the graduated bottle is filled with additional double distilled H
2
O to reach the
final volume of 1000.0 ml. Continue the carbogen bubbling until the pH of the buffer solution reaches
7.4.
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