Media formula

TAP (Tris-Acetate Phosphate)
To make one liter of TAP:


 * 10 mL 2M Tris-acetate (pH 7.0)
 * 10 mL Phosphate buffer (pH 7.0)
 * 10 mL nutrient stock
 * 10 mL trace metals (10x dilution)
 * Bring to 1 liter with distilled water

Variations

 * For plates add 15g agar per liter (1.5%)
 * For TAP-sulfur substitute MgCl2 for the MgSO4 in the nutrient stock and omit trace metals

2M Tris-acetate stock

 * 242 g Tris base
 * Dissolve in 600 mL water while titrating to pH 7.0 with glacial acetic acid.
 * Bring to 1 liter

Phosphate buffer

 * 10.8 g K2HPO4
 * 5.6 g KH2PO4
 * Bring to 1 liter

Nutrient stock

 * 40 g NH4Cl
 * 10 g MgSO4·7H2O
 * 5 g CaCl2·2H2O
 * Bring to 1 liter

Trace metals
Making this is a complex proceedure so (unless you think you will be using a lot of it) there are a couple of alternatives.
 * Make a soil extract by shaking a few grams of soil in water
 * Use a small amount of Job's plant food spike
 * Use tap water rather than distilled.

This makes a liter of full-strength, enough for 1000 L of TAP. Dilute an aliquot tenfold and use the diluted stock in making TAP


 * 1) Dissolve 50g of acid free EDTA in 250 ml of ddH2O. Heat to dissolve.
 * 2) Dissolve the following, one by one, in order, heating to approximately 100° in 550 ml ddH2O.
 * 3) *11.40 g BO3H3
 * 4) *22.00 g ZnSO4·7H2O
 * 5) *5.06 g MnCl2·4H2O
 * 6) *4.99 g FeSO4·7H2O
 * 7) *1.61 g CoCl2·6H2O
 * 8) *1.57 g CuSO4·5H2O
 * 9) *1.1 g Mo7O24(NH4)6·4H2O (Ammonium Molybdate)
 * 10) Mix the two solutions (1 & 2) together. The resulting solution should be blue-green.
 * 11) Heat to 100°C. Cool slightly, but don't let the temperature drop below 80°-90°.
 * 12) Adjust pH to 6.5 to 6.8 with 20% KOH (approximately 83 ml). Don't let the temperature drop below 70° until after the pH is adjusted.
 * 13) Make up to 1L and let stand in a 2L Erlenmeyer flask, stoppered loosly. The color should change from green to purple over a period of days.
 * 14) Remove the rust-coloured precipitate by filtering, with suction, through 3 layers of Whatman #1 filter paper in a Buchner funnel. Repeat until no more precipitate is seen on the filter paper
 * 15) Store in a brown bottle at 4°.

Links
Recipe taken from: http://www.chlamyteach.info/growthmedium.html