263. SINGULISPHAERA MEDIUM

N-acetylglucosamine 1.0 g

KH₂PO₄ 0.1 g

Hutner’s basal salts medium 20.0 ml

Peptone 0.1 g

Yeast extract 0.1 g

Agar-agar (Difco) 18.0 g

Distilled water 1000.0 ml

Adjust pH to 5.8

Hutner’s basal salts medium:

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) 50.0 ml

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

pH 7.2 (adjust with KOH or H₂SO₄).

Adjust volume to 1000.0 ml with distilled water.

Metal 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.

Autoclave at 121°C 15 min.

 

Updated 1.02.2024