Phosphate buffer
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Purpose
Phosphate buffer is a multi-purpose buffer used for many molecular biology procedures.
How to make phosphate buffer
Source: Sörensen; in Hayat, 1986.
0.2 M monobasic stock
- 15.6 g sodium phosphate monobasic dihydrate (NaH2PO4·2H2O, MW 155.99 g/mol), or 13.9 g of the anhydrous form
- 500 mL dH2O
0.2 M dibasic stock
- 53.65 g sodium phosphate dibasic heptahydrate (Na2HPO4·7H2O, MW 268.07 g/mol), or 28.4 g of the anhydrous form
- 1 L dH2O
0.1 M sodium phosphate buffer
To make 600mL 0.1 M sodium phosphate buffer, combine the following quantities of each stock and add dH2O to bring up to 600 mL:
monobasic (mL) | dibasic (mL) | pH |
280.5 | 19.5 | 5.7 |
276 | 24 | 5.8 |
270 | 30 | 5.9 |
263.1 | 36.9 | 6.0 |
255 | 45 | 6.1 |
244.5 | 55.5 | 6.2 |
232.5 | 67.5 | 6.3 |
220.5 | 79.5 | 6.4 |
205.5 | 94.5 | 6.5 |
187.5 | 112.5 | 6.6 |
169.5 | 130.5 | 6.7 |
153 | 147 | 6.8 |
135 | 165 | 6.9 |
117 | 183 | 7.0 |
99 | 201 | 7.1 |
84 | 216 | 7.2 |
69 | 231 | 7.3 |
57 | 243 | 7.4 |
48 | 252 | 7.5 |
39 | 261 | 7.6 |
31.5 | 271.5 | 7.7 |
25.5 | 274.5 | 7.8 |
21 | 279 | 7.9 |
15.9 | 284.1 | 8.0 |
Use
Safety
- Are there any safety considerations for this material?
References
- Any papers or websites that are relevant to the use of this material