ES DK medium

Fe stock
To 350ml ddH20 add

1) Fe(NH4)2 (SO4) 2. 6H20 0.351g

11) Na2 EDTA 0.3565g

Make final vol. up to 500ml with ddH20

2) P2 stock

To 300ml ddH2O. add

a) Na2 EDTA 0.400g

b) FeCl3. 6H2O 0.126g
c) H3 B03 0.456g
d) Mn SO4. H20 0.055g
e) Zn SO4. H20 0.0088g
f) Co SO4. 7H2O 0.00192g

Make final volume up to 400ml with ddH2O

*Autoclave. Both stock solutions at 12°C 20min cool the solution

immediately after
Autoc. To avoid precipitation. Left 24 hours befour use.
*Stored at 4°C

To make ES working stock

a) NaNO3 1.400g
b) Na=glycerophosphate 0.200g
c) P2 stock 100ml
d) Fe stock 100ml

make up volume to 400ml with ddH2O autoclave. Store at 4°C into

Teflon bottle

To make the ES DK medium add 9.3ml of ES working stock into 1liter

filtered autoclave seawater

Use f/2 vit sol.

Flask autoclave 20minit
  Seawater was filtered through 0.2µm capsul filter and autoclave at
121°C for 30minit cool rapidly after autoclaving.
 Keep the seawater for at least 2 days befour use ( for gases diffuse
into the liquid)
 Check the pH,( 7.8-82 is desirable)

1Liter ES DK= 5ml ES stock + Soil ext 3ml + Vit sol 0.5ml