Methanosarcina DPB Medium

[所属分类:培养基配方] [发布时间:2021-5-26] [发布人:网站管理员2] [阅读次数:] [返回]

Methanosarcina DPB Medium

山东拓普生物工程有限公司 培养基配方 http://www.topbiol.com

Composition per 1001.0mL:

Sodium acetate·3H 2 O....................................................................4.1g
NH 4 Cl ...........................................................................................1.4g
K 2 HPO 4 .........................................................................................1.3g
KH 2 PO 4 .........................................................................................1.3g
MgSO 4 ..........................................................................................0.5g
NaCl..............................................................................................0.5g
L -Cysteine·HCl·H 2 O...................................................................0.27g
CaCl 2 ·2H 2 O ................................................................................0.06g
Fe(NH 4 ) 2 (SO 4 ) 2 ..........................................................................0.01g
Antifoam C..............................................................................10.0mL
Trace elements solution...........................................................10.0mL
Vitamin solution.......................................................................10.0mL
Na 2 S·9H 2 O solution...................................................................1.0mL
pH 6.8 ± 0.2 at 25°C
Source: Antifoam C is available from Sigma Chemical Co.
Trace Elements Solution:
Composition per liter:
MgSO 4 ·5H 2 O................................................................................3.0g
Nitrilotriacetic acid.......................................................................1.5g
NaCl..............................................................................................1.0g
MnSO 4 ·2H 2 O................................................................................0.5g
NaS 4 ·SeO 3 ·5H 2 O ..........................................................................0.3g
NiCl 2 ·6H 2 O.................................................................................0.25g
CoSO 4 ·7H 2 O...............................................................................0.18g
ZnSO 4 ·7H 2 O...............................................................................0.18g
CaCl 2 ·7H 2 O ..................................................................................0.1g
FeSO 4 ·7H 2 O..................................................................................0.1g
KAI(SO 4 ) 2 ·12H 2 O ......................................................................0.02g
CuSO 4 ·5H 2 O...............................................................................0.01g
H 3 BO 3 .........................................................................................0.01g
Na 2 MoO 4 ·2H 2 O..........................................................................0.01g
Preparation of Trace Elements Solution:  Add nitrilotriacetic
acid to 500.0mL of distilled/deionized water. Adjust pH to 6.5 with
KOH. Add remaining components. Mix thoroughly. Add distilled/de-
ionized water to 1.0L. Adjust pH to 6.8.
Vitamin Solution:
Composition per liter:
Calcium  DL -pantothenate..............................................................5.0g
Vitamin B 12 ...................................................................................0.1g
Pyridoxine·HCl........................................................................10.0mg
p-Aminobenzoic acid.................................................................5.0mg
Lipoic acid.................................................................................5.0mg
Nicotinic acid.............................................................................5.0mg
Riboflavin..................................................................................5.0mg
Thiamine·HCl............................................................................5.0mg
Biotin.........................................................................................2.0mg
Folic acid ...................................................................................2.0mg
Preparation of Vitamin Solution:  Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly.
Na 2 S·9H 2 O Solution:
Composition per 100.0mL:
Na 2 S·9H 2 O..................................................................................25.0g
Preparation of Na 2 S·9H 2 O Solution:  Bring 100.0mL of dis-
tilled/deionized water to boiling. Cool to room temperature while
sparging with 100%N 2 . Add Na 2 S·9H 2 O to the 100.0mL of anaerobic
water. Mix thoroughly. Distribute into serum bottles fitted with butyl
rubber stoppers and aluminum seals. Do not grease stoppers. Auto-
clave for 20 min at 15 psi pressure–121°C.
Preparation of Medium: Add components, except Na 2 S·9H 2 O so-
lution, to distilled/deionized water and bring volume to 1.0L. Mix thor-
oughly. Adjust pH to 6.8 with concentrated HCl. Distribute into tubes
or flasks. Autoclave for 15 min at 15 psi pressure–121°C. Cool medi-
um while sparging with 100% N 2 . Prior to inoculation, aseptically and
anaerobically add 1.0mL of sterile Na 2 S·9H 2 O solution per liter of me-
dium. Repeat the addition of 1.0mL of sterile Na 2 S·9H 2 O solution per
liter of medium every 48 hr during growth.
Use:  For the cultivation of Methanosarcina species.