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
13.9 g sodium phosphate monobasic dihydrate (Na2HPO4·2H2O, MW 155.99 g/mol) 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
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