TITLE: K/2 Ian / K-ET
AUTHORS: Ian Probert

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">Modified from K medium by Ian Probert</div><div class = "text-block">Reference </div><div class = "text-block">Keller, M.D., Selvin, R.C., Claus, W. & Guillard, R.R.L. 1987. Media for the culture of oceanic ultraphytoplankton. J. Phycol. 23:633–8.</div></div>

[STEPS]
?. ABCD1Quantity
Compound
Stock solution (sterile)
Final conc. in K medium
20.5ml
NaNO3
48.9542g/litre H2O
288µM
30.5ml
NH4Cl *
0.535g/litre H2O
5µM
40.5ml
KH2PO4
4.8992g/litre H2O
18µM
50.5ml
FeEDTA solution
(see recipe below)
(see below)
60.5ml
Trace metal solution
(see recipe below)
(see below)
71.0ml
f/2 vitamin solution
(see recipe below)
(see below)
* optional
ABCD1Quantity
Compound
Stock solution (sterile)
Final conc. in K medium
20.5ml
NaNO3
48.9542g/litre H2O
288µM
30.5ml
NH4Cl *
0.535g/litre H2O
5µM
40.5ml
KH2PO4
4.8992g/litre H2O
18µM
50.5ml
FeEDTA solution
(see recipe below)
(see below)
60.5ml
Trace metal solution
(see recipe below)
(see below)
71.0ml
f/2 vitamin solution
(see recipe below)
(see below)
?. [FeEDTA solution]
To 950ml distilled H2O add:    Quantity
   Compound
   Stock solution
   Final conc. in K medium
     4.3g
   (Na)FeEDTA
   -
   5.85µM
    Make up to 1 litre with milliQ H2O, sterilize (filter 0.22µm) and store in fridge.  ABCD1Quantity
Compound
Stock solution
Final conc. in K medium
24.3g
(Na)FeEDTA
-
5.85µM
ABCD1Quantity
Compound
Stock solution
Final conc. in K medium
24.3g
(Na)FeEDTA
-
5.85µM
?. [Trace metal solution]
To 950ml distilled H2O add:     Quantity
   Compound
   Stock solution
   Final conc. in K medium
     37.22g
   Na2EDTA.2H2O
   -
   50µM
     1.0ml
   CuSO4.5H2O
   2.497g/litre H2O
   0.005µM
     1.0ml
   Na2MoO4.2H2O
   7.2585g/litre H2O
   0.015µM
     1.0ml
   ZnSO4.7H2O
   23.0g/litre H2O
   0.004µM
     1.0ml
   CoSO4.7H2O
   14.055g/litre H2O
   0.025µM
     1.0ml
   MnCl2.4H2O
   178.11g/litre H2O
   0.45µM
     1.0ml
   H2SeO3
   1.29g/litre H2O
   0.005µM
     1.0ml
   NiCl2.6H20
   1.49 g/litre H2O
   0.00314µM
     Make up to 1 litre with milliQ H2O, sterilize (filter 0.22µm) and store in fridge.  ABCD1Quantity
Compound
Stock solution
Final conc. in K medium
237.22g
Na2EDTA.2H2O
-
50µM
31.0ml
CuSO4.5H2O
2.497g/litre H2O
0.005µM
41.0ml
Na2MoO4.2H2O
7.2585g/litre H2O
0.015µM
51.0ml
ZnSO4.7H2O
23.0g/litre H2O
0.004µM
61.0ml
CoSO4.7H2O
14.055g/litre H2O
0.025µM
71.0ml
MnCl2.4H2O
178.11g/litre H2O
0.45µM
81.0ml
H2SeO3
1.29g/litre H2O
0.005µM
91.0ml
NiCl2.6H20
1.49 g/litre H2O
0.00314µM
ABCD1Quantity
Compound
Stock solution
Final conc. in K medium
237.22g
Na2EDTA.2H2O
-
50µM
31.0ml
CuSO4.5H2O
2.497g/litre H2O
0.005µM
41.0ml
Na2MoO4.2H2O
7.2585g/litre H2O
0.015µM
51.0ml
ZnSO4.7H2O
23.0g/litre H2O
0.004µM
61.0ml
CoSO4.7H2O
14.055g/litre H2O
0.025µM
71.0ml
MnCl2.4H2O
178.11g/litre H2O
0.45µM
81.0ml
H2SeO3
1.29g/litre H2O
0.005µM
91.0ml
NiCl2.6H20
1.49 g/litre H2O
0.00314µM
?. [f/2 Vitamin solution]
To 950ml distilled H2O add     Quantity
   Compound
   Stock solution
   Final conc. in K medium
     1.0ml
   Vit. B12 (cyanocobalamin)
   0.5g/litre H2O
   0.37nM
     1.0ml
   Biotin
   5.0mg/litre H2O
   2.0nM
     100.0mg
   Thiamine HCl
   -
   0.3µM
     Make up to 1 litre with milliQ H2O, filter sterilize into plastic vials and store in freezer.  ABCD1Quantity
Compound
Stock solution
Final conc. in K medium
21.0ml
Vit. B12 (cyanocobalamin)
0.5g/litre H2O
0.37nM
31.0ml
Biotin
5.0mg/litre H2O
2.0nM
4100.0mg
Thiamine HCl
-
0.3µM
ABCD1Quantity
Compound
Stock solution
Final conc. in K medium
21.0ml
Vit. B12 (cyanocobalamin)
0.5g/litre H2O
0.37nM
31.0ml
Biotin
5.0mg/litre H2O
2.0nM
4100.0mg
Thiamine HCl
-
0.3µM
?. [Sterilization of medium]
Optional: Heat to 80°C for 2 hours and leave to cool – this should kill most organisms but should not chemically modify the medium too muchFilter sterilize through 0.22µm filters (e.g. Millipore Steritop units) into sterile (autoclaved) polycarbonate bottles.
?. [For K-ET]
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