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ATCC medium: 1551 Methanomicrobium medium

中國(guó)微生物菌種查詢(xún)網(wǎng) / 2017-01-06 12:38:22

ATCC medium: 1551 Methanomicrobium medium

Rumen fluid (clarified)................300.0 ml

K2HPO4 ...................................0.3 g

KH2PO4 ...................................0.3 g

(NH4)2SO4.................................0.3 g

NaCl.....................................0.6 g

MgSO4 . 7H2O .............................0.13 g

CaCl2 . 2H2O .............................8.0 mg

Wolfe's Mineral Solution (see below)....10.0 ml

Wolfe's Vitamin Solution (see below)....10.0 ml

FeSO4 . 7H2O .............................2.0 mg

NaHCO3 ...................................2.0 g

Yeast Extract (BD 212750)................1.0 g

Trypticase Peptone (BD 211921)...........1.0 g

Fatty Acid Mixture (see below)..........20.0 ml

Resazurin (0.1%).........................1.0 ml

L-Cysteine . HCl.........................0.5 g

Na2S . 9H2O ..............................0.5 g

Distilled water........................660.0 ml

Adjust final pH to 6.5. Tube under 80% N2, 20% CO2 in aluminum seal tubes.

Fatty Acid Mixture:

Valeric acid.......................0.7 ml

Isovaleric acid....................0.7 ml

alpha-Methylbutyric acid...........0.5 ml

Isobutyric acid....................0.5 ml

Distilled water....................1.0 L

Wolfe's Vitamin Solution:

Available from ATCC as a sterile ready-to-use liquid (Vitamin Supplement, catalog no. MD-VS).

Biotin.............................2.0 mg

Folic acid.........................2.0 mg

Pyridoxine hydrochloride..........10.0 mg

Thiamine . HCl.....................5.0 mg

Riboflavin.........................5.0 mg

Nicotinic acid.....................5.0 mg

Calcium D-(+)-pantothenate.........5.0 mg

Vitamin B12........................0.1 mg

p-Aminobenzoic acid................5.0 mg

Thioctic acid......................5.0 mg

Distilled water....................1.0 L

2 Wolfe's Mineral Solution:

Available from ATCC as a sterile ready-to-use liquid (Trace Mineral Supplement, catalog no. MD-TMS.)

Nitrilotriacetic acid..............1.5 g

MgSO4 . 7H2O .......................3.0 g

MnSO4 . H2O ........................0.5 g

NaCl...............................1.0 g

FeSO4 . 7H2O .......................0.1 g

CoCl2 . 6H2O .......................0.1 g

CaCl2 ..............................0.1 g

ZnSO4 . 7H2O .......................0.1 g

CuSO4 . 5H2O .......................0.01 g

AlK(SO4)2 . 12H2O...................0.01 g

H3BO3 ..............................0.01 g

Na2MoO4 . 2H2O......................0.01 g

Distilled water....................1.0 L

Add nitrilotriacetic acid to approximately 500 ml of water and adjust to pH 6.5 with KOH to dissolve the compound. Bring volume to 1.0 L with remaining water and add remaining compounds one at a time.

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