Difference between revisions of "Part:BBa K389423:Experience"
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'''Characterization tests''' | '''Characterization tests''' | ||
| − | Cultivation was done by induction with | + | Cultivation was done by induction with acetosyringone at 50 µM. Controls were not induced sensitivity tuner devices as well as induced and not induced native system ([https://parts.igem.org/Part:BBa_K389015 K389015]; without tuning elements). Induction was done upon inoculation. Measuring point for amplification factor calculation was OD 1.0. |
'''Results''' | '''Results''' | ||
| − | Three sensitivity tuned | + | Three sensitivity tuned ''vir''-gene sensing systems were obtained: [https://parts.igem.org/Part:BBa_K389421 K389421], [https://parts.igem.org/Part:BBa_K389422 K389422] and [https://parts.igem.org/Part:BBa_K389423 K389423] distinguishing by the amplification level of ''luc'' transcription. |
| − | [[Image:Bielefeld amp factor.png|400px|thumb|center| '''Figure 1: Amplification factor of induced, 50 µM | + | [[Image:Bielefeld amp factor.png|400px|thumb|center| '''Figure 1: Amplification factor of induced, 50 µM acetosyringone (red) and not induced (green) modified sensitivity tuner [https://parts.igem.org/Part:BBa_K389421 K389421], [https://parts.igem.org/Part:BBa_K389422 K389422] and [https://parts.igem.org/Part:BBa_K389423 K389423], standard deviation shown.''']] |
The amplification factor was received by apply [https://parts.igem.org/Part:BBa_K389015 K389015] as reference. Amplification calculation was done by normalizing relative luminescence units emitted from luciferase per OD. | The amplification factor was received by apply [https://parts.igem.org/Part:BBa_K389015 K389015] as reference. Amplification calculation was done by normalizing relative luminescence units emitted from luciferase per OD. | ||
| − | Output-signal amplification is in the induced contructs (red) [https://parts.igem.org/Part:BBa_K389422 K389422] and [https://parts.igem.org/Part:BBa_K389423 K389423] 100 and respectively 200 fold higher than in not induced controls (green). An exception is K389422 were induced and not | + | Output-signal amplification is in the induced contructs (red) [https://parts.igem.org/Part:BBa_K389422 K389422] and [https://parts.igem.org/Part:BBa_K389423 K389423] 100 and respectively 200 fold higher than in not induced controls (green). An exception is K389422 were induced and not induced system revealed analog results. Corresponding to data of iGEM Team, Cambridge 2009, K389423 (originated from [https://parts.igem.org/Part:BBa_I746390 I746390]) shows the highest amplification rate of all tested sensitivity tuners. Our results indicate to higher amplification rate of [https://parts.igem.org/Part:BBa_K389421 K389421] than [https://parts.igem.org/Part:BBa_K389422 K389422] of 100 fold under induced conditions. The controls also show high basal transcription rates. |
| − | Because there is small difference in induced and not induced system | + | Because there is small difference visible in induced and not induced system and basal transcription rates are high, we assume that the sensitivity tuning constructs are not well applicable for luciferase measurements. |
| − | For further theory click [http://2010.igem.org/Team:Bielefeld-Germany/Project/Theory#Read_out_system | + | For further theory click [http://2010.igem.org/Team:Bielefeld-Germany/Project/Theory#Read_out_system read out system] |
| − | For test results click [http://2010.igem.org/Team:Bielefeld-Germany/Results/Tests#BBa_K389421.2C_BBa_K389422.2C_BBa_K389423:_Sensitivity_Tuner_amlified_Vir-test_system | + | For test results click [http://2010.igem.org/Team:Bielefeld-Germany/Results/Tests#BBa_K389421.2C_BBa_K389422.2C_BBa_K389423:_Sensitivity_Tuner_amlified_Vir-test_system sensitivity tuner amlified Vir-test system] |
===User Reviews=== | ===User Reviews=== | ||
Latest revision as of 00:29, 28 October 2010
This experience page is provided so that any user may enter their experience using this part.
Please enter
how you used this part and how it worked out.
Applications of BBa_K389423
Characterization tests
Cultivation was done by induction with acetosyringone at 50 µM. Controls were not induced sensitivity tuner devices as well as induced and not induced native system (K389015; without tuning elements). Induction was done upon inoculation. Measuring point for amplification factor calculation was OD 1.0.
Results
Three sensitivity tuned vir-gene sensing systems were obtained: K389421, K389422 and K389423 distinguishing by the amplification level of luc transcription.
The amplification factor was received by apply K389015 as reference. Amplification calculation was done by normalizing relative luminescence units emitted from luciferase per OD. Output-signal amplification is in the induced contructs (red) K389422 and K389423 100 and respectively 200 fold higher than in not induced controls (green). An exception is K389422 were induced and not induced system revealed analog results. Corresponding to data of iGEM Team, Cambridge 2009, K389423 (originated from I746390) shows the highest amplification rate of all tested sensitivity tuners. Our results indicate to higher amplification rate of K389421 than K389422 of 100 fold under induced conditions. The controls also show high basal transcription rates.
Because there is small difference visible in induced and not induced system and basal transcription rates are high, we assume that the sensitivity tuning constructs are not well applicable for luciferase measurements.
For further theory click [http://2010.igem.org/Team:Bielefeld-Germany/Project/Theory#Read_out_system read out system]
For test results click [http://2010.igem.org/Team:Bielefeld-Germany/Results/Tests#BBa_K389421.2C_BBa_K389422.2C_BBa_K389423:_Sensitivity_Tuner_amlified_Vir-test_system sensitivity tuner amlified Vir-test system]
User Reviews
UNIQ9d27b0f905a0fed5-partinfo-00000000-QINU UNIQ9d27b0f905a0fed5-partinfo-00000001-QINU