Team:DTU-Denmark/Notebook/9 August 2013

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{{:Team:DTU-Denmark/Templates/StartPage|9 August 2013}}
{{:Team:DTU-Denmark/Templates/StartPage|9 August 2013}}
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Navigate to the [[Team:DTU-Denmark/Notebook/8_August_2013|Previous]] or the [[Team:DTU-Denmark/Notebook/10_August_2013|Next]] Entry
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=lab 208=
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=Lab 208=
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==Main purpose==
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==Who was in the lab==
==Who was in the lab==
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Kristian
==Procedure==
==Procedure==
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===Gel purification===
===Gel purification===
Gel purified AMO with USER endings and Nir1 extraction fragment made in yesterdays PCR.
Gel purified AMO with USER endings and Nir1 extraction fragment made in yesterdays PCR.
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==Results==
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=Lab 115=
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==Conclusion==
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=lab 115=
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==Main purpose==
==Main purpose==
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Run experiment 2: "Measuring the production of N<sub>2</sub>O from Nitrite NO<sub>2</sub><sup>-<sup>
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Run [[Team:DTU-Denmark/Experiment2|Experiment 2]] anaerobically in order to characterize the behavior of ''E.coli''.
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anaerobically in order to characterize the behavior of ''E.coli''.
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==Who was in the lab==
==Who was in the lab==
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Following the protocol "Experiment 2: Measuring production of N<sub>2</sub>O from Nitrite NO<sub>2</sub><sup>-</sup> anaerobically"
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Following the protocol [[Team:DTU-Denmark/Methods/Determining_concentration_of_nitrogen_compounds/Experiment_2|Experiment 2]]
Changing the steps :
Changing the steps :
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7. Then added 3 ml more after 2 hours and the OD was 0.1046. The OD after 1 hour was 0.1340. Afterwards we added glycerol 1g in 10 ml of water and then added in our growing culture. Then after 40 min the OD was 0.2737. (Setup wavelength 600 nm).
7. Then added 3 ml more after 2 hours and the OD was 0.1046. The OD after 1 hour was 0.1340. Afterwards we added glycerol 1g in 10 ml of water and then added in our growing culture. Then after 40 min the OD was 0.2737. (Setup wavelength 600 nm).
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8. We didn't cool down the centrifuge because the experiment had to be done in 37 degrees.
 
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11. The volume was 200 ml instead of 100 ml.
 
12. The OD was measured 0.1324 instead of 0.3.
12. The OD was measured 0.1324 instead of 0.3.
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15. 10 minutes of N<sub>2</sub> saturation instead of 5 minutes.
 
19. Add 0.5 ml of nitrite solution and continue by adding 0.5 ml after 10 minutes then we took 2.2 ml of sample, we added then 1 ml and we saw a slow response after 6 minutes. Afterwards when there was any change in the curve we spiked with 1 ml of nitrite. We continued as there was no response by adding 1 ml more and then by adding 2 ml more. We took 2 ml for sample in the end and another 2 ml for OD measurement where OD=0.1350. One hour after we took 2 ml of sample and then we spiked with 1 ml of NO<sub>2</sub>. There was no response after 12 minutes. Then we spiked with 0.5 ml of N<sub>2</sub>O and we show a big response.
19. Add 0.5 ml of nitrite solution and continue by adding 0.5 ml after 10 minutes then we took 2.2 ml of sample, we added then 1 ml and we saw a slow response after 6 minutes. Afterwards when there was any change in the curve we spiked with 1 ml of nitrite. We continued as there was no response by adding 1 ml more and then by adding 2 ml more. We took 2 ml for sample in the end and another 2 ml for OD measurement where OD=0.1350. One hour after we took 2 ml of sample and then we spiked with 1 ml of NO<sub>2</sub>. There was no response after 12 minutes. Then we spiked with 0.5 ml of N<sub>2</sub>O and we show a big response.
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Point before spiking with NO<sub>2</sub> at the end
Point before spiking with NO<sub>2</sub> at the end
measurement 1.03 mg/L (x2 dilution)- 0.5 mg/L (x4 dilution)
measurement 1.03 mg/L (x2 dilution)- 0.5 mg/L (x4 dilution)
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===N<sub>2</sub>O measurement===
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[[File:DTU-Experiment-2-results.png|600px]]
==Conclusion==
==Conclusion==
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The ''E.coli'' didn't grow as usual and the problem might be the minimal medium.
The ''E.coli'' didn't grow as usual and the problem might be the minimal medium.
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Concerning the N<sub>2</sub>O sensor, after adding for third time 1 ml of nitrite the response was slow and started to get stabilized after 6 minutes.
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When we continue adding more nitrite there was no other response.
Navigate to the [[Team:DTU-Denmark/Notebook/8_August_2013|Previous]] or the [[Team:DTU-Denmark/Notebook/10_August_2013|Next]] Entry
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{{:Team:DTU-Denmark/Templates/EndPage}}
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Latest revision as of 11:55, 4 October 2013

9 August 2013

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Contents

Lab 208

Who was in the lab

Kristian

Procedure

Gel purification

Gel purified AMO with USER endings and Nir1 extraction fragment made in yesterdays PCR.

Lab 115

Main purpose

Run Experiment 2 anaerobically in order to characterize the behavior of E.coli.

Who was in the lab

Ariadni, Helen, Natalia

Procedure

Adjusting the temperature at 37 degrees and calibrating the probes as described in Appendix 5.


Following the protocol Experiment 2

Changing the steps :

6. 3 ml of the overnight culture (from glycerol stock) in 200 ml of medium with OD=0.0317.

7. Then added 3 ml more after 2 hours and the OD was 0.1046. The OD after 1 hour was 0.1340. Afterwards we added glycerol 1g in 10 ml of water and then added in our growing culture. Then after 40 min the OD was 0.2737. (Setup wavelength 600 nm).

12. The OD was measured 0.1324 instead of 0.3.

19. Add 0.5 ml of nitrite solution and continue by adding 0.5 ml after 10 minutes then we took 2.2 ml of sample, we added then 1 ml and we saw a slow response after 6 minutes. Afterwards when there was any change in the curve we spiked with 1 ml of nitrite. We continued as there was no response by adding 1 ml more and then by adding 2 ml more. We took 2 ml for sample in the end and another 2 ml for OD measurement where OD=0.1350. One hour after we took 2 ml of sample and then we spiked with 1 ml of NO2. There was no response after 12 minutes. Then we spiked with 0.5 ml of N2O and we show a big response.

Results

Colorimetric results

Ammonium

Measuring range 2-75 mg/L NH4-N

start point- signal <2 mg/L

middle point- signal 2 mg/L

end point- signal <2 mg/L


Nitrate

Measuring range 1-25 mg/L NO3-N

start point- signal <1 mg/L

middle point- signal <1 mg/L but visible pinker

end point- signal <1 mg/L but much visible pinker


Nitrite

Measuring range 0.02-1 mg/L NO2-N

start point- signal <0.02 mg/L

middle point- signal 0.29 mg/L

end point- signal 0.70 mg/L after X2 dilution


Point before spiking with NO2 at the end measurement 1.03 mg/L (x2 dilution)- 0.5 mg/L (x4 dilution)


N2O measurement

DTU-Experiment-2-results.png

Conclusion

The E.coli didn't grow as usual and the problem might be the minimal medium. Concerning the N2O sensor, after adding for third time 1 ml of nitrite the response was slow and started to get stabilized after 6 minutes. When we continue adding more nitrite there was no other response.

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