Product category Bacteria
Product type Extremophile
Strain designation 別名 Nb-101
Type strain No
Product format 提供形式 Frozen
General
Preceptrol No
Handling information
Medium 培養(yǎng)基 ATCC Medium 480: Nitrobacter medium 203
Temperature 培養(yǎng)溫度 26°C
Atmosphere 需氧情況 Aerobic
IncubationWith shakingHandling procedure1. Allow vial to thaw. Transfer the entire contents into 5.0 ml of #480 broth contained in a large test tube (20 X 150 mm). 2. Incubate tube at 26oC, static, in the dark, and in a slanted position to increase surface area.
3. Monitor culture growth daily by assaying for the disappearance of NO2 in the culture medium. Withdraw 0.1 ml from the culture and place in a small test tube. Add one drop of NO2 Reagent A followed by one drop of NO2 Reagent B, mixing after each addition. Pink color development occurs within 10 minutes and is directly related to amount of NO2 present. Initial tests will probably be dark magenta reflecting the 20 mM NO2 concentration of medium #480; but as culture grows and NO2 is oxidized, color will become magenta, pink and finally clear. This process can take from 3 to 20 days depending on the viability of the stock.
4. When all NO2 has been oxidized, inoculate a 250 ml flask containing 100 ml of medium #480 with the 5.0 ml tube culture. Wrap the flask in foil to protect from light and incubate at 26oC with gentle shaking. Monitor NO2 concentration daily.
5. When NO2 is again depleted in the culture medium, either transfer the culture to fresh media or feed the culture by adding additional NO2. A 10% inoculum is recommended for transfers. To feed cultures, aseptically add sterile NaNO2 solution, returning NO2 concentrations to approximately 10 mM (3.5 ml of 2% NaNO2 per 100 ml culture). Cells should be harvested or sub-cultured to fresh media when cultures can utilize 10 mM NO2 in less than 24 hours and cell numbers are greater than 10 cells per field at 1000X magnification.
6. Cells may be harvested by centrifugation. For long-term storage, 1.0 ml aliquots of concentrated cells can be frozen with glycerol as the cryoprotectant at a final concentration of 10%. Frozen vial should be stored at
?70
Deposited asNitrobacter sp.DepositorsSW Watson
Chain of custodyATCC <-- SW Watson <-- H. Funk <-- M. Alexander (Nitrobacter agilis)
Cross references GenBank L35506 Nitrobacter winogradskyi (ATCC 25381) 16S ribosomal RNA (16S rRNA) gene.
