Difference between revisions of "Team:Gifu/More/"

 
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<center><font size="20" face="Century">Inter Lab Study</font></center><br><br>
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<center><font size="20" face="Century">InterLab Study</font></center><br><br>
  
 
<font size="6" face="Century">Equipment</font>  
 
<font size="6" face="Century">Equipment</font>  
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<p>&nbsp;&nbsp;We constructed 5 plasmids [positive control, negative control, device1~3] by using BioBricks and then transformed to <i>E. coli</i> K-12 JM109. We checked them by colony direct PCR and plasmids sequence. We cultivated correct bacterias by shaking until OD660:0.5. Cultivated mediums were　transferred to 96-well plate by 100mL. We read the absorbance.</p>
 
<p>&nbsp;&nbsp;We constructed 5 plasmids [positive control, negative control, device1~3] by using BioBricks and then transformed to <i>E. coli</i> K-12 JM109. We checked them by colony direct PCR and plasmids sequence. We cultivated correct bacterias by shaking until OD660:0.5. Cultivated mediums were　transferred to 96-well plate by 100mL. We read the absorbance.</p>
 
<br>
 
<br>
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&nbsp;&nbsp;Setting of plate reader is following:<br></font>
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&nbsp;&nbsp;Setting of plate reader is following:<br>
 
<img src="https://static.igem.org/mediawiki/2015/2/20/Set_safari_gify.png" border="0" width="395" height="220">
 
<img src="https://static.igem.org/mediawiki/2015/2/20/Set_safari_gify.png" border="0" width="395" height="220">
 
<br><br><br><br>
 
<br><br><br><br>
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</ul>
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<b>Sequence data</b><br>
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<br>
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・device 1 : J23101+ I13504<br>
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<img src="https://static.igem.org/mediawiki/2015/6/6b/Sequence_data1.png" width="800"><br><br>
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<br>
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・device 2 : J23106+ I13504<br>
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<img src="https://static.igem.org/mediawiki/2015/6/60/Sequence_data2.png" width="800"><br><br>
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<br>
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・device 3 : J23117+ I13504<br>
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<img src="https://static.igem.org/mediawiki/2015/c/c7/Sequence_data3.png" width="800"><br><br>
  
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<br>These sequence datas are correct.<br></font>
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</ul>
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<br><br>
 
<font size="6" face="Century">Result</font>  
 
<font size="6" face="Century">Result</font>  
 
<hr width="100%" ><br>
 
<hr width="100%" ><br>
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<img src="https://static.igem.org/mediawiki/2015/a/a7/Interlab_data.jpg" width="800"><br><br>
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<img src="https://static.igem.org/mediawiki/2015/a/a7/Interlab_data.jpg" width="830"><br><br><br><br>
  
 
<li> <font size="5" face="Century"> Value<br></font> </li>
 
<li> <font size="5" face="Century"> Value<br></font> </li>
 
<font size="4" face="Century">
 
<font size="4" face="Century">
 
<p>Relative absorbance values for positive control are shown below. <br></p>
 
<p>Relative absorbance values for positive control are shown below. <br></p>
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<br>
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<br><img src="https://static.igem.org/mediawiki/2015/d/de/Interlab_relative.jpg" width="350"></img><br>
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<br>
 
<img src="https://static.igem.org/mediawiki/2015/e/e4/GURAHU-gifu.png" border="0" width="572" height="364"> <br><br>
 
<img src="https://static.igem.org/mediawiki/2015/e/e4/GURAHU-gifu.png" border="0" width="572" height="364"> <br><br>
 
                       FIG1 Relative absorbance values <br></font>
 
                       FIG1 Relative absorbance values <br></font>
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               20A : Device1 (J23101 + I13504)<br>
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               20K : Device1 (J23101 + I13504)<br>
 
               22A : Device2 (J23106 + I13504)<br>
 
               22A : Device2 (J23106 + I13504)<br>
 
               22K : Device3 (J23117 + I13504)<br><br><br>
 
               22K : Device3 (J23117 + I13504)<br><br><br>
  
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<br><img src="https://static.igem.org/mediawiki/2015/0/05/Gifu_interlab_posicon1.jpeg" width="400px"></img><br>
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</ul>
 
</ul>
 
  <br><br>
 
  <br><br>

Latest revision as of 17:26, 18 September 2015


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InterLab Study


Equipment

  • BioSHAKER TA-12R
    • use for shaking culture



  • Safire ART-NR:F129013
    • use for measurement of absorbance




Method of Measurement

      We constructed 5 plasmids [positive control, negative control, device1~3] by using BioBricks and then transformed to E. coli K-12 JM109. We checked them by colony direct PCR and plasmids sequence. We cultivated correct bacterias by shaking until OD660:0.5. Cultivated mediums were　transferred to 96-well plate by 100mL. We read the absorbance.


      Setting of plate reader is following:




    Sequence data

    ・device 1 : J23101+ I13504



    ・device 2 : J23106+ I13504



    ・device 3 : J23117+ I13504



    These sequence datas are correct.


Result

  • Definition

    •   We expressed absorbance as relative value for　positive control.
    This expression is following:
      Sample : Absorbance in sample
      NCav : Average of absorbance in negative control
      PCav : Average of absorbance in positive control






  • Value

    • Relative absorbance values for positive control are shown below.








    FIG1 Relative absorbance values
    20K : Device1 (J23101 + I13504)
    22A : Device2 (J23106 + I13504)
    22K : Device3 (J23117 + I13504)




Discussion

      The value of device1 is much higher than that of device2 and 3. This is why each device is different from promoter sequence, especially -10 and -35 region.



    Promoter sequence in device1 resembles consensus sequence. On the other hand, device2 and 3 is not similar than device1. This is how each GFP expression is different.









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