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

 
(18 intermediate revisions by the same user not shown)
Line 69: Line 69:
 
<div id="main">
 
<div id="main">
 
<blockquote>
 
<blockquote>
−
<center><font size="20" face="Century">Inter Lab Study</font></center><br><br>
+
<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>  
Line 90: Line 90:
 
<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>
−
&nbsp;&nbsp;Setting of plate reader is following:<br></font>
+
&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>
−
</ul>
+
<b>Sequence data</b><br>
 +
<br>
 +
・device 1 : J23101+ I13504<br>
 +
<img src="https://static.igem.org/mediawiki/2015/6/6b/Sequence_data1.png" width="800"><br><br>
 +
<br>
 +
・device 2 : J23106+ I13504<br>
 +
<img src="https://static.igem.org/mediawiki/2015/6/60/Sequence_data2.png" width="800"><br><br>
 +
<br>
 +
・device 3 : J23117+ I13504<br>
 +
<img src="https://static.igem.org/mediawiki/2015/c/c7/Sequence_data3.png" width="800"><br><br>
  
 +
<br>These sequence datas are correct.<br></font>
 +
</ul>
 +
<br><br>
 
<font size="6" face="Century">Result</font>  
 
<font size="6" face="Century">Result</font>  
 
<hr width="100%" ><br>
 
<hr width="100%" ><br>
Line 107: Line 119:
 
<br>
 
<br>
 
<br>
 
<br>
−
<img src="https://static.igem.org/mediawiki/2015/6/6d/Gifu_interlab_rawdata.jpeg" width="700px"></img><br>
+
<img src="https://static.igem.org/mediawiki/2015/a/a7/Interlab_data.jpg" width="830"><br><br><br><br>
−
<br>
+
−
<br><img src="https://static.igem.org/mediawiki/2015/9/99/Gifu_interlab_well.jpeg" width="700px"></img><br>
+
−
<br>
+
−
<br><img src="https://static.igem.org/mediawiki/2015/a/a6/Gifu_interlab_negacon.jpeg" width="700px"></img><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>
 +
<br>
 +
<br><img src="https://static.igem.org/mediawiki/2015/d/de/Interlab_relative.jpg" width="350"></img><br>
 +
<br>
 +
<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>
−
               20A : Device1 (J23101 + I13504)<br>
+
               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>
 +
 +
 
</ul>
 
</ul>
 
  <br><br>
 
  <br><br>

Latest revision as of 17:26, 18 September 2015


https://static.igem.org/mediawiki/2015/d/d8/Logo_HOME.png https://static.igem.org/mediawiki/2015/a/a4/Logo_PROJECT.png https://static.igem.org/mediawiki/2015/archive/b/b9/20150720104417%21Logo_TEAM.png https://static.igem.org/mediawiki/2015/8/86/Logo_NOTE.png https://static.igem.org/mediawiki/2015/8/87/Logo_RESULT.png https://static.igem.org/mediawiki/2015/2/28/Logo_MORE.png


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.









  MENU
  1. InterLab Study   
  2. SAFETY 
  3. Policy & Practice