280. PLANCTOMYCES MEDIUM

KH₂PO₄ 0.1 g

(NH₄)₂SO₄ 0.2 g

MgSO₄ × 7 H₂O 0.1 g

CaCl₂ × 6 H₂O 0.02 g

NaCl 0.01 g

N-acetylglucosamine 1.0 g

Glucose 0.5 g

Yeast e×tract 0.2 g

Hutner's basal salts (see below) 1.0 ml

Staley's vitamin solution (see below) 1.0 ml

Phytagel (for solid medium) 8.0 g

Distilled water 1000.0 ml

Adjust pH to 5.8-6.5

Autoclave at 121°C 15 min. Add filter-sterilized vitamin solution

Hutner's basal salts:

Nitrilotriacetic acid (NTA) 10.0 g

MgSO₄ × 7 H₂O 29.7 g

CaCl₂ × 2 H₂O 3.34 g

Na₂MoO₄ × 2 H₂O 12.67 mg

FeSO₄ × 7 H₂O 99.0 mg

Metal salt solution "44" (see below) 50 ml

Dissolve NTA first by neutralizing with KOH, then add other salts.

pH 7.2 (ajust with KOH or H₂SO₄).

Ajust volume to 1000.0 ml with distilled water.

Metal salt solution "44":

Na-EDTA 250.0 mg

ZnSO₄ × 7 H₂O 1095.0 mg

FeSO₄ × 7 H₂O 500.0 mg

MnSO₄ × 7 H₂O 154.0 mg

CuSO₄ × 5 H₂O 39.0 mg

Co(NO₃)₂ × 6 H₂O 24.8 mg

Na₂B₄O₇ × 10 H₂O 17.7 mg

Distilled water 1000.0 ml

Dissolve Na-EDTA and add a few drops of concentrated H₂SO₄ to retard precipitation of heavy metal ions.

Staley’s vitamin solution:

Vitamin B₁₂ 0.10 mg

Biotin 2.0 mg

Thiamine-HCl × 2 H₂O 5.0 mg

Ca-pantothenate 5.0 mg

Folic acid 2.0 mg

Riboflavin 5.0 mg

Nicotinamide 5.0 mg

p-Aminobenzoic acid 5.0 mg

Pyridoxine-HCl 10.0 mg

Distilled water 1000.0 ml

Store in the dark and cold (5°C).