嗜热自养甲烷杆菌噬菌体Methanothermobacter marburgensis phage BioVector NTCC质粒载体菌种细胞基因保藏中心

嗜热自养甲烷杆菌噬菌体Methanothermobacter marburgensis phage BioVector NTCC质粒载体菌种细胞基因保藏中心
Product Sheet
Species:
Methanobacterium phage psiM1
Taxonomy:
Order: Caudovirales
Family: Siphoviridae
Type species of the genus: Psimunalikevirus
Isolate:
psi M2
NTCC No.:
P12792
History:
<- T. Leisinger
Isolated from:
anaerobic sludge digester
Date of sampling:
before 1990
Genbank accession numbers:
complete genome: AF065411
Genomics:
spontaneous deletion variant of the wild-type phage psi M1
Morphology:
icosahedral head, long non-contractile tail
Further information:
Molecular analysis (7672). Transduction experiments (7671, 7673).
Risk group:
1
Phage propagation
Host bacterium:
Methanothermobacter marburgensis Wasserfallen et al. 2000
We strongly recommend to use only Methanothermobacter marburgensis for propagation.
Cultivation conditions:
METHANOBACTERIUM MEDIUM , anaerobic, 65°C

METHANOBACTERIUM MEDIUM
KH2PO4 0.50 g
MgSO4 x 7 H2O 0.40 g
NaCl 0.40 g
NH4Cl 0.40 g
CaCl2 x 2 H2O 0.05 g
FeSO4 x 7 H2O solution (0.1% w/v in 0.1 N H2SO4) 2.00 ml
Trace element solution SL-10 (see medium 320) 1.00 ml
Yeast extract (OXOID) 1.00 g
Na-acetate 1.00 g
Na-formate 2.00 g
Sludge fluid (see below) 50.00 ml
Fatty acid mixture (see below) 20.00 ml
Na-resazurin solution (0.1% w/v) 0.50 ml
NaHCO3 4.00 g
L-Cysteine-HCl x H2O 0.50 g
Na2S x 9 H2O 0.50 g
Distilled water 930.00 ml
Dissolve ingredients except bicarbonate, cysteine and sulfide. Sparge medium with 80%
H2 and 20% CO2 gas mixture for 30 – 45 min to make it anoxic. Add and dissolve
bicarbonate, then dispense medium under 80% H2 and 20% CO2 gas atmosphere into
anoxic Hungate-type tubes and autoclave. Add cysteine and sulfide from sterile anoxic
stock solutions prepared under 100% N2 gas. Prior to use check pH of complete medium
and adjust to 6.8 - 7.0, if necessary.
Note: After growth has started and the culture is becoming turbid add sterile 80% H2
and 20% CO2 gas mixture to 0.5 - 1 bar overpressure.
Sludge fluid:
Add 0.4% yeast extract to sludge from an anaerobic digester, and after gassing with
nitrogen gas for a few minutes incubate it at 37C for 24 hours. Then centrifuge the
sludge at 13000 g and autoclave the resulting, clear supernatant in screw-capped
vessels under nitrogen gas. The sludge fluid can be stored at room temperature in the
dark.
 Fatty acid mixture:
Valeric acid 25.00 g
Isovaleric acid 25.00 g
2-Methylbutyric acid 25.00 g
Isobutyric acid 25.00 g
Distilled water 1000.00 ml
Adjust pH to 7.5 with conc. NaOH.
Phage propagation:
Supply, storage and propagation of phages
Revitalisation of Vaccuum-Dried Phages



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【Website网址】 http://www.biovector.net