MediaDive DSMZ
Content
Media Solutions Ingredients Strains Steps Gas
Search
Medium finder Solution finder Taxonomy search Isolation sources
Tools
Medium builder NEW Unit converter Compare media Compare solutions Prediction SOON
DSMZ

DSMZ 169b . LEADBETTERA MEDIUM

What is this page?
Equipment needed: Autoclave Filter Gassing station Hungate tubes
Final gas composition: 100 % N2 read more
Final pH: 7.5
Select a strain (optional):

ml
Compound Amount Unit
Solution A 952 ml
Solution B 20 ml
Solution C 20 ml
Solution D 1 ml
Solution E 10 ml
1 Dissolve ingredients of solution A (except cysteine) and sparge medium with 100% N2 gas for 30 - 45 min to make it anoxic. Add cysteine and adjust pH 7.2 with KOH, then distribute under the same gas atmosphere in anoxic Hungate-type tubes or serum vials and autoclave. Solution B is autoclaved separately under 80% N2 and 20% CO2 gas atmosphere, solution C is autoclaved under 100% N2 gas atmosphere and solutions D and E are sterilized by filtration under 100% N2 gas atmosphere. To complete the medium appropriate amounts of solutions B, C, D and E are added to the sterile solution A in the sequence as indicated. Final pH of the complete medium should be 7.5.

Solution A #

952
Compound Amount Unit
CaCl2 x 2 H2O 0.04 g
MgSO4 x 7 H2O 0.50 g
Yeast extract
(OXOID) 		4.00 g
D-Glucose 4.00 g
Trace element solution SL-10 1.00 ml
Selenite-tungstate solution 1.00 ml
Sodium resazurin
(0.1% w/v) 		0.50 ml
L-Cysteine HCl x H2O 0.50 g
Distilled water 950.00 ml

Solution B #

20
Compound Amount Unit
NaHCO3 1 g
Distilled water 20 ml

Solution C #

20
Compound Amount Unit
K2HPO4
(0.5 M) 		16.04 ml
KH2PO4
(0.5 M) 		3.96 ml
1 Combine 16.04 ml of 0.5 M K2HPO4 with 3.96 ml of 0.5 M KH2PO4 stock solution to reach a pH of around 7.4.

Solution D #

1
Compound Amount Unit
Seven vitamins solution 1 ml

Solution E #

10
Compound Amount Unit
DL-Dithiothreitol 0.15 g
Distilled water 10.00 ml

Trace element solution SL-10 #

1000
Compound Amount Unit
HCl
(25%) 		10.0 ml
FeCl2 x 4 H2O 1.5 g
ZnCl2 70.0 mg
MnCl2 x 4 H2O 100.0 mg
H3BO3 6.0 mg
CoCl2 x 6 H2O 190.0 mg
CuCl2 x 2 H2O 2.0 mg
NiCl2 x 6 H2O 24.0 mg
Na2MoO4 x 2 H2O 36.0 mg
Distilled water 990.0 ml
1 First dissolve FeCl2 in the HCl, then dilute in water, add and dissolve the other salts. Finally make up to 1000.00 ml.

Selenite-tungstate solution #

1000
Compound Amount Unit
NaOH 0.5 g
Na2SeO3 x 5 H2O 3.0 mg
Na2WO4 x 2 H2O 4.0 mg
Distilled water 1000.0 ml

Seven vitamins solution #

1000
Compound Amount Unit
Vitamin B12 100 mg
p-Aminobenzoic acid 80 mg
D-(+)-biotin 20 mg
Nicotinic acid 200 mg
Calcium pantothenate 100 mg
Pyridoxine hydrochloride 300 mg
Thiamine-HCl x 2 H2O 200 mg
Distilled water 1000 ml