Difference between revisions of "Team:NU Kazakhstan/InterLab"

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<h3> InterLab Study </h3>
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<p>This page is used by the judges to evaluate your team for the <a href="https://2018.igem.org/Judging/Medals">medal criterion</a> or <a href="https://2018.igem.org/Judging/Awards"> award listed below</a>. </p>
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Every year, the Measurement Committee tries to analyze the causes of difference in values of fluorescence measurements obtained in laboratories around the year. Particularly in this year, the main goal of the Fifth International InterLaboratory Measurement Study is to investigate if the normalization of fluorescence measurements to the absolute cell count can contribute to the reduction of lab-to-lab variability in results. Our team, NU_Kazakhstan 2018, used the Varioskan LUX Multimode Microplate Reader to measure the fluorescence of DH 5 𝛼 cells transformed with the GFP inserted in pSB1C3 plasmid with different promoters (BBa_J364000, BBa_J364001, BBa_J364002,  BBa_J364007, BBa_J364008, BBa_J364009, plus negative BBa_R0040 and positive BBa_I20270 controls).
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<h1>InterLab</h1>
 
<h1>InterLab</h1>
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<h3>Bronze Medal Criterion #4</h3>
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<h3></h3>
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<p><b>Standard Tracks:</b> Participate in the Interlab Measurement Study and/or obtain new, high quality experimental characterization data for an existing BioBrick Part or Device and enter this information on that part's Main Page in the Registry. The part that you are characterizing must NOT be from a 2018 part number range.
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<p><b></b> Every year, the Measurement Committee tries to analyze the causes of difference in values of fluorescence measurements obtained in laboratories around the year. Particularly in this year, the main goal of the Fifth International InterLaboratory Measurement Study is to investigate if the normalization of fluorescence measurements to the absolute cell count can contribute to the reduction of lab-to-lab variability in results. Our team, NU_Kazakhstan 2018, used the Varioskan LUX Multimode Microplate Reader to measure the fluorescence of DH 5 𝛼 cells transformed with the GFP inserted in pSB1C3 plasmid with different promoters (BBa_J364000, BBa_J364001, BBa_J364002,  BBa_J364007, BBa_J364008, BBa_J364009, plus negative BBa_R0040 and positive BBa_I20270 controls).
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<h3><b>Protocol</b></h3>
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For teams participating in the <a href="https://2018.igem.org/Measurement/InterLab">InterLab study</a>, all work must be shown on this page.  
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We strictly followed the instructions from the protocol provided by iGEM https://static.igem.org/mediawiki/2018/0/09/2018_InterLab_Plate_Reader_Protocol.pdf
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Plate reader configuration:
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<h4><i>Photometric</i></h4>
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Wavelength: 600 nm
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Bandwidth: 5 nm
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Measurement time: 100 ms
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<h4><i>Fluorometric</i></h4>
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Excitation Wavelength: 485 nm
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Emission Wavelength: 530 nm
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Excitation bandwidth: 12 nm
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Fluorescence reading: top optics
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Measurement time: 100 ms
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<h3><b>Results</b></h3>
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OD600 reference point
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<h4>Conclusion</h4>
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The data from measurements shows that test device 4 (BBa_J364007) has the promoter with the highest fluorescence among the rest of the provided promoters. Its maximum value is 11.84 net fluorescein a.u. after 6 hours of incubation. The weakest promoter was found to be the test device 3 (max. value 0.48 net fluorescein a.u. at 6 hours). Samples transformed with test devices 1,5, and 6 have about similar results (approx. 2 net fluorescein a.u.). The fluorescence of samples transformed with test device 2 is slightly higher than these three. Its maximum value of fluorescence is 5.53 net fluorescein a.u.
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</p>
 
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Revision as of 12:57, 25 July 2018

Bioremediation of Sour Crude Oil Waste using Cyanobacteria




InterLab

Every year, the Measurement Committee tries to analyze the causes of difference in values of fluorescence measurements obtained in laboratories around the year. Particularly in this year, the main goal of the Fifth International InterLaboratory Measurement Study is to investigate if the normalization of fluorescence measurements to the absolute cell count can contribute to the reduction of lab-to-lab variability in results. Our team, NU_Kazakhstan 2018, used the Varioskan LUX Multimode Microplate Reader to measure the fluorescence of DH 5 𝛼 cells transformed with the GFP inserted in pSB1C3 plasmid with different promoters (BBa_J364000, BBa_J364001, BBa_J364002, BBa_J364007, BBa_J364008, BBa_J364009, plus negative BBa_R0040 and positive BBa_I20270 controls).

Protocol

We strictly followed the instructions from the protocol provided by iGEM https://static.igem.org/mediawiki/2018/0/09/2018_InterLab_Plate_Reader_Protocol.pdf Plate reader configuration:

Photometric

Wavelength: 600 nm Bandwidth: 5 nm Measurement time: 100 ms

Fluorometric

Excitation Wavelength: 485 nm Emission Wavelength: 530 nm Excitation bandwidth: 12 nm Fluorescence reading: top optics Measurement time: 100 ms

Results

OD600 reference point

Conclusion

The data from measurements shows that test device 4 (BBa_J364007) has the promoter with the highest fluorescence among the rest of the provided promoters. Its maximum value is 11.84 net fluorescein a.u. after 6 hours of incubation. The weakest promoter was found to be the test device 3 (max. value 0.48 net fluorescein a.u. at 6 hours). Samples transformed with test devices 1,5, and 6 have about similar results (approx. 2 net fluorescein a.u.). The fluorescence of samples transformed with test device 2 is slightly higher than these three. Its maximum value of fluorescence is 5.53 net fluorescein a.u.