Team:DTU-Denmark/Methods/Bioreactor
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# Take 100 mL as the first sample t=0 for both colorimetric analysis and nitrous oxide measurement, from each of the 3 vessels. | # Take 100 mL as the first sample t=0 for both colorimetric analysis and nitrous oxide measurement, from each of the 3 vessels. | ||
# After 10 hours take another 100 ml of sample to measure nitrous oxide and make the colorimetric analysis, from each of the 3 vessels. | # After 10 hours take another 100 ml of sample to measure nitrous oxide and make the colorimetric analysis, from each of the 3 vessels. | ||
+ | # After taking the samples pour them in 200 ml sealed vessels with syringes in order to keep the anaerobic conditions. | ||
#To finish gathering data, make the colorimetric measurements for nitrite and ammonium for each of the samples collected by using the protocols [[Team:DTU-Denmark/Methods/Nitrite colorimetric measurements|Nitrite colorimetric measurements]] and [[Team:DTU-Denmark/Methods/Ammonium colorimetric measurements|Ammonium colorimetric measurements]], respectively. | #To finish gathering data, make the colorimetric measurements for nitrite and ammonium for each of the samples collected by using the protocols [[Team:DTU-Denmark/Methods/Nitrite colorimetric measurements|Nitrite colorimetric measurements]] and [[Team:DTU-Denmark/Methods/Ammonium colorimetric measurements|Ammonium colorimetric measurements]], respectively. |
Revision as of 13:40, 1 October 2013
Bioreactor
Procedure
In this anaerobic experiment we add nitrite in our cultures with the Nir transformed cells, in order to test how nitrite is converted to nitrous oxide.
- Make an overnight culture with the Nir transformation cells and then pour 100 ml in each of the 3 replicates with 800 ml DM minimal medium.
- Set the bio-reactor at 37oC.
- Add 0.8 ml of 0.916 M ammonium chloride solution in each of the three vessels.
- Add 8 ml of 50mM of nitrite stock solution to each of the three vessels.
- Take 100 mL as the first sample t=0 for both colorimetric analysis and nitrous oxide measurement, from each of the 3 vessels.
- After 10 hours take another 100 ml of sample to measure nitrous oxide and make the colorimetric analysis, from each of the 3 vessels.
- After taking the samples pour them in 200 ml sealed vessels with syringes in order to keep the anaerobic conditions.
- To finish gathering data, make the colorimetric measurements for nitrite and ammonium for each of the samples collected by using the protocols Nitrite colorimetric measurements and Ammonium colorimetric measurements, respectively.