Phosphate buffer

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Contents

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
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