IGEM08 How to measure phosphate

Reagent I

 * Sulfuric acid (H2SO4)
 * Potassium antimonyl tartrate solution
 * Ammonium molybdate solution

Reagent II

 * Ascorbic acid (0.1M): Dissolve 1.76 g ascorbic acid in 100 mL distilled water

Volume ratio between reagents

 * 10X Sulfuric acid (H2SO4)
 * 1X Potassium antimonyl tartrate solution
 * 3X Ammonium molybdate solution
 * 6X ascorbic acid

Reaction procedure

 * sample 1mL (1000 uL)
 * reagent I (112 uL)
 * reagent II (48uL)


 * STEP 1: mix sample with reagent I
 * STEP 2: vortex the mixture
 * STEP 3: add reagent II (reaction starts!)

Measure procedure

 * 10 mins reaction time (each sample MUST have equal reaction time)
 * BECKMAN DU-600 at 880nm (圖資大樓6F共儀)
 * Turn VIS 'ON' for at least 5 mins
 * Set parameter 880nm as 'ON'
 * Read blank
 * Read samples
 * Turn VIS 'OFF'

Other methods

 * A continuous spectrophotometric assay for inorganic phosphate and for measuring phosphate release kinetics in biological systems, Proc Natl Acad Sci U S A. 1992