20
WIRE MYOGRAPH SYSTEM - 620M
- USER GUIDE
APPENDIX 1
APPENDIX 1 - BUFFER RECIPES
Physiological Saline Solution (PSS)
1x PSS:
Chemical
Mol.Wt
mM
g/0.5L
g/L
g/2L
g/4L
NaCl
(58.45)
130
3.799
7.598
15.20
30.39
KCl
(74.557)
4.7
0.175
0.35
0.70
1.40
KH
2
PO
4
(136.09)
1.18
0.08
0.16
0.32
0.64
MgSO
4
7H
2
O
(246.498)
1.17
0.145
0.29
0.58
1.16
NaHCO
3
(84.01)
14.9
0.625
1.25
2.50
5.00
Glucose
(180.16)
5.5
0.5
1.00
2.00
4.00
EDTA
(380)
0.026
0.005
0.01
0.02
0.04
CaCl
2
(110.99)
1.6
0.8mL
1.6mL
3.2mL
6.4mL
(1.0 M solution)
1. Make a 1.0M solution of CaCl
2
(110.99) in double-distilled H
2
O. Filter-sterilize the calcium solution through a 0.22 μm filter.
The sterilized solution can be stored in the refrigerator for up to 3 months.
2. Dissolve all the chemicals except the CaCl
2
in approximately 80% of the desired final volume of double distilled H
2
O while
being constantly stirred. For example, if 1 litre of PSS is to be made, then dissolve all the chemicals in 800mL of double
distilled H
2
O.
3. Add the appropriate volume of 1.0M CaCl
2
for the total volume of PSS being made (for example, 1.6mL of 1.0M CaCl
2
for 1
litre of buffer). Continue to stir the PSS while the CaCl
2
is being added.
4. Bring the solution up to the final volume with double-distilled H
2
O. Continue to stir the solution until the EDTA is fully
dissolved. This takes about 15 minutes at room temperature.
5. Aerate the solution with carbogen (95% O
2
+ 5% CO
2
) for about 20 minutes.
25x Concentrated PSS:
Chemical
Mol.Wt
mM
g/0.5L
g/L
g/2L
g/4L
NaCl
(58.45)
3250
94.98
189.96
379.92
759.84
KCl
(74.557)
117.5
4.375
8.75
17.5
35.0
KH
2
PO
4
(136.09)
29.5
2.0
4.0
8.0
16.0
MgSO
4
7H
2
O
(246.498)
29.25
3.625
7.25
14.5
29.0
NaHCO
3
(84.01)
14.9
0.625
1.25
2.50
5.00
Glucose
(180.16)
5.5
0.5
1.00
2.00
4.00
EDTA
(380)
0.65
0.125
0.25
0.50
1.0
CaCl
2
(110.99)
40
20mL
40mL
80mL
160mL
(1.0 M solution)
