Difference between revisions of "Part:BBa K1456002:Experience"

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This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
 
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how you used this part and how it worked out.
 
  
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==WESTERN BLOTTİNG==
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[[File:ATOMS gpx results 11.jpg|400px|]]
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<li>Sadly, the results achieved did not fit the ideal Blotting results.
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==WESTERN BLOTTİNG==
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[[File:ATOMS gpx results 13.jpg|400px|]]
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<li>Selenium was necessary for the production of the selenocysteine amino acid present in GPx but was not present in the medium of the culture. It was expected to have observed a result such as the figure on the left through adding Na2SeO3 to the cell line. Expression was seen; there was a directly proportional increase in GPx production in respect to selenoite increase(100 uM-200 uM-500 uM-1000 uM).<br/>
  
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[[File:ATOMS gpx results 15.jpg|400px|]]
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<li>The bands acquired through the Western Blotting were fitting the expected results. The cell lines, that now had their transfections verified, were purfied for the preparation of GPx assays.<br/>
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[[File:ATOMS gpx results 16.jpg|400px|]]
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<li>The amounts of protein acquired were as the charts above and the these data were taken into consideration in the preparation of the assays.<br/>
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[[File:ATOMS gpx results 17.jpg|400px|]]
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<li>A functional assay of GPx was done through NADPH to measure GPx activity. The same amount of genes were transferred to all samples but varying amount of Na2SeO3 was prepared in the samples, with 2 samples containing different Na2SeO3 concentrations and one control group. The GPx gene was not transfected to the control group. From these 3 groups, the assay of the control group gave the lowest yield as expected; the sample of 500 nM Na2SeO3 produced the mid-result. The sample of 1000 nM Na2SeO3 yielded the highest activity results in the assay. These results prove that the GPx production is directly proportional to the amount of added Na2Se2O3 and that the GPx enzyme was produced functionally.
  
 
===Applications of BBa_K1456002===
 
===Applications of BBa_K1456002===
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===ATOMS TURKİYE 2014===
 
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                        https://static.igem.org/mediawiki/2014/0/0e/ATOMS_gpx_results_16.jpg
 
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The amounts of protein acquired were as the charts above and the these data were taken into consideration in the preparation of the assays.
 
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                        https://static.igem.org/mediawiki/2014/d/df/ATOMS_gpx_results_17.jpg
 
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A '''functional assay''' of GPx was done '''through NADPH to measure GPx activity'''. The same amount of genes were transferred to all samples but varying amount of Na2SeO3 was prepared in the samples, with 2 samples containing different Na2SeO3 concentrations and one control group. The GPx gene was not transfected to the control group. From these 3 groups, the assay of the control group gave the lowest yield as expected; the sample of 500 nM Na2SeO3 produced the mid-result. The sample of 1000 nM Na2SeO3 yielded the highest activity results in the assay. These results prove that the GPx production is directly proportional to the amount of added Na2Se2O3 and that the GPx enzyme was produced functionally.
 
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== Characterisation ==
 
  
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== ATOMS TURKİYE 2014 ==
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===User Reviews===
 
===User Reviews===
 
<!-- DON'T DELETE --><partinfo>BBa_K1456002 StartReviews</partinfo>
 
<!-- DON'T DELETE --><partinfo>BBa_K1456002 StartReviews</partinfo>

Latest revision as of 14:09, 21 October 2014


WESTERN BLOTTİNG

ATOMS gpx results 11.jpg

  • Sadly, the results achieved did not fit the ideal Blotting results.

    WESTERN BLOTTİNG

    ATOMS gpx results 13.jpg

  • Selenium was necessary for the production of the selenocysteine amino acid present in GPx but was not present in the medium of the culture. It was expected to have observed a result such as the figure on the left through adding Na2SeO3 to the cell line. Expression was seen; there was a directly proportional increase in GPx production in respect to selenoite increase(100 uM-200 uM-500 uM-1000 uM).
    ATOMS gpx results 15.jpg
  • The bands acquired through the Western Blotting were fitting the expected results. The cell lines, that now had their transfections verified, were purfied for the preparation of GPx assays.
    ATOMS gpx results 16.jpg
  • The amounts of protein acquired were as the charts above and the these data were taken into consideration in the preparation of the assays.
    ATOMS gpx results 17.jpg
  • A functional assay of GPx was done through NADPH to measure GPx activity. The same amount of genes were transferred to all samples but varying amount of Na2SeO3 was prepared in the samples, with 2 samples containing different Na2SeO3 concentrations and one control group. The GPx gene was not transfected to the control group. From these 3 groups, the assay of the control group gave the lowest yield as expected; the sample of 500 nM Na2SeO3 produced the mid-result. The sample of 1000 nM Na2SeO3 yielded the highest activity results in the assay. These results prove that the GPx production is directly proportional to the amount of added Na2Se2O3 and that the GPx enzyme was produced functionally.

    Applications of BBa_K1456002

    ATOMS TURKİYE 2014

    User Reviews

    UNIQ7109a07ef33a7d9e-partinfo-00000000-QINU

    UNIQ7109a07ef33a7d9e-partinfo-00000001-QINU