User manual

55
User Manual vers. 1.4 Chapter 7
7.3 Buffer Recipes
7.3.1 Physiological Saline Solution (PSS)
1x PSS (1000ml):
Solution 1.
Chemical MW (g/mol) Conc. (mmol/l) Conc. (g/l)
NaCl 58.44 118.99 6.954
KCl 74.56 4.69 0.350
MgSO
4
- 7H
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
- 2H
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
a) Dissolve the chemicals in approximately 100ml double distilled H
2
O as three
individual solutions as described in the table above. Gently heat solution 3 to
dissolve the EDTA.
b) Solution 1 is added to a graduated bottle and the bottle is lled with double
distilled H
2
O to a nal volume of 500ml.
c) Solution 3 is added to the graduated bottle, which afterwards is lled with
additional double distilled H
2
O to a nal volume of about 850ml.
d) Aerate the solution with carbogen (95% O
2
+ 5% CO
2
) for about 20 minutes.
e) Solution 2 is added and the graduated bottle is lled with additional double
distilled H
2
O to reach the nal volume of 1000 ml. Continue the carbogen
bubbling until the pH of the buffer solution reaches 7.4.