Previous Page Table of Contents Next Page


Appendix 1
THE CULTURE MEDIA AND THEIR PREPARATION

1. Allen-Nelson “Miquel seawater”

Solution A
KNO320.2 g/100 ml.H20
Solution B
Na2HP04.12H20  4.0 g
Cacl2.6H20  4.0 g
HCl conc.  2.0 g
FeCl3 (melted)  2.0 ml
Distilled water80.0 ml

Add 2 ml. solution A to 1 liter seawater and 1 ml. of solution B to 1 liter of seawater.

II. Conwy Medium (Walne, 1974)

A.Sodium Nitrate100 g
EDTA, disodium salt45 g
Boric Acid33.6 g
Sodium Phosphate, monobasic  20 g
Ferric Chloride, 6-hydrate1.3 g
Manganous Chloride, 4-hydrate0.36 g
Trace Metal Solution*   1 ml
Vitamin Mix**100 ml
Distilled water (to make)1000 ml
(Note: use 1 ml Conwy medium/liter of seawater)
* Trace Metal Stock Solution
Zinc Chloride2.1 g
Cobalt Chloride, 6-hydrate2.1 g
Ammonium Molybdate, 4-hydrate2.1 g
Copper Sulfate,2.0 g
Distilled Water100 ml.
(Note: acidify with IN HCl until solution is clear)
** Vitamin Mix
Vitamin B1210.0 mg
Vitamin B120.0 mg
Distilled water200 ml

III. TMRL Enrichment (Liao & Huang, 1970)

KNO3100 g
Na2HPO4.12H2010 g
FeCl3.6H203 g
Na2SiO3.9H201 g
Distilled water1000 ml
(Note: Use 1 ml TMRL/liter of seawater)

IV. Enrichment for Outdoor Cultures

16–20–012 g
Urea 4612 g
21–0–0100 g
(Note: add above nutrients to 1 ton seawater)

V. Modified F Medium (modified from Guillard & Ryther, 1962)

A.N-P Stock (500x)
Sodium nitrate42.07 g
Sodium Phosphate, monobasic5.00 g
Distilled water (to make)1 liter
B.Sodium Metasilicate Stock (500x)
Sodium metasilicate15.0 g
Distilled water (to make)1 liter
C.Ferric Chloride Stock (500x)1.45 g
Distilled water (to make)1 liter
D.EDTA Stock (1000x)
EDTA, Disodium salt10.00 g
Distilled water (to make)1 liter
E.Vitamin stock (1000x) 
Thiamine HCl0.2 g
B1210.0 ml*
Biotin10.0 ml**
* B12 stock (primary)
-B120.1 g
Distilled
water 1 liter
** Biotin primary stock
Biotin0.1 g
Distilled water1 liter
F.Trace Metal Stock (1000x)
TM primary stock A1 ml
TM primary stock B1 ml
TM primary stock C1 ml
TM primary stock D1 ml
Distilled water (to make)1 liter
1) TM primary stock A solution
Copper sulphate, 5-hydrate1.96 g
Zinc sulphate, 7-hydrate4.40 g
Distilled water (to make)100 ml
2) TM primary stock B solution
Sodium molybdate, 2-hydrate1.26 g
or
(Ammonium molybdate, 4-hydrate) 0.907 g
Distilled water (to make)100 ml
3) TM primary stock C solution
Manganous Chloride36.00 g
Distilled water (to make)100 ml
4) TM primary stock D solution
Cobalt Chloride2.00 g
Distilled water (to make)100 ml

NOTE: Dispense 2 ml each of solutions A, B, & C/liter of seawater

Dispense 1 ml each of solutions D, E, & F/liter of seawater.


Previous Page Top of Page Next Page