CASTENHOLZ TYE MEDIUM, Modified

Complete Medium:

Castenholz Salts 2X (see below) 500.0 ml

TYE Solution (see below) 100.0 ml

Agar (if needed) 15.0 g

Distilled deionized water 400.0 ml

Combine aseptically. Adjust for final pH 7.6 If agar is used, add it to the water component.

Autoclave at 121°C for 15 minutes.

Castenholz Salts, 2X:

Nitrilotriacetic acid 0.2 g

Nitsch s Trace Elements (see below) 2.0 ml

FeCl3 solution (0.03%) 2.0 ml

CaSO4 . 2H2O 0.12 g

MgSO4 . 7H2O 0.2 g

NaCl 0.016 g

KNO3 0.21 g

NaNO3 1.4 g

Na2HPO4 0.22 g

Distilled water 1.0 L

Adjust pH to 8.2. May be stored at 2-5°C for one year. Filter-sterilize before use.

Nitsch s Trace Elements:

H2SO4 0.5 ml

MnSO4 2.2 g

ZnSO4 0.5 g

H3BO3 0.5 g

CuSO4 0.016 g

Na2MoO4 0.025 g

CoCl2 . 6H2O 0.046 g

Distilled water 1.0 L

TYE Solution:

Tryptone (BD 211705) 20.0 g

Yeast extract 20.0 g

Distilled water 1.0 L