MS low iron

Submitted by kdorfman on Mon, 12/19/2011 - 21:12

**Murashige & Skoog 1µM iron medium

MS 10x micronutrients is 100 µM FeSO4, so MS complete is 10 µM

FeSO4 stock is 10 mM, which is 10,000x 1 µM

To make plates low-iron MS
start with mL ddH2O (~ 60% final volume)
add: mL 10X macronutrients
add: mL boric acid 1000X
add: mL cobalt chloride 10,000X
add: mL cupric sulfate 10,000X
add: mL ferrous sulfate 10 mM
add: mL KI 10,000X
add: mL manganese sulfate 1000X
add: mL moybdic acid 10,000X
add: mL zinc sulfate 1000X
add: g MES, pH to 5.7 with KOH
bring to mL final volume
add: g bacto- or phyto-agar
autoclave for minutes

ingredient 750 1000 1200 1500 mL
number of plates 25 33 40 50
ddH2O* 450 600 600 960 mL
10X macronutrients 75 100 120 150 mL
boric acid 1000x 0.75 1.0 1.2 1.5 mL
Cobalt chloride 10,000x 0.075 0.1 0.12 0.15 mL
cupric sulfate 10,000x 0.075 0.1 0.12 0.15 mL
ferrous sulfate 10,000x 0.075 0.1 0.12 0.15 mL
KI 10,000x 0.075 0.1 0.12 0.15 mL
manganese sulfate 1000x 0.75 1.0 1.2 1.5 mL
molybdic acid 10,000x x 0.075 0.1 0.12 0.15 mL
zinc sulfate 1000x 0.75 1.0 1.2 1.5 mL
MES 0.375 0.5 0.6 0.75 g

*Add the other salt mixtures to the water to prevent precipitation

pH to 5.7 with 1M KOH (initial pH = ~4.6) (needs ~570 drops/L)

Bring to volume with distilled water

ingredient 750 1000 1200 1500 mL
bacto or phyto agar 7.5 10 12 15 g

autoclave with stir bar in flask or bottle

Stir until cool enough to handle

Pour 30 mL per plate (use the deep ones)