Team:AUC TURKEY/Results/Functional Essay

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 +
  <w:LsdException Locked="false" Priority="67" Name="Medium Grid 1"/>
 +
  <w:LsdException Locked="false" Priority="68" Name="Medium Grid 2"/>
 +
  <w:LsdException Locked="false" Priority="69" Name="Medium Grid 3"/>
 +
  <w:LsdException Locked="false" Priority="70" Name="Dark List"/>
 +
  <w:LsdException Locked="false" Priority="71" Name="Colorful Shading"/>
 +
  <w:LsdException Locked="false" Priority="72" Name="Colorful List"/>
 +
  <w:LsdException Locked="false" Priority="73" Name="Colorful Grid"/>
 +
  <w:LsdException Locked="false" Priority="60" Name="Light Shading Accent 1"/>
 +
  <w:LsdException Locked="false" Priority="61" Name="Light List Accent 1"/>
 +
  <w:LsdException Locked="false" Priority="62" Name="Light Grid Accent 1"/>
 +
  <w:LsdException Locked="false" Priority="63" Name="Medium Shading 1 Accent 1"/>
 +
  <w:LsdException Locked="false" Priority="64" Name="Medium Shading 2 Accent 1"/>
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  <w:LsdException Locked="false" Priority="65" Name="Medium List 1 Accent 1"/>
 +
  <w:LsdException Locked="false" SemiHidden="true" Name="Revision"/>
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  <w:LsdException Locked="false" Priority="34" QFormat="true"
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  Name="List Paragraph"/>
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  <w:LsdException Locked="false" Priority="29" QFormat="true" Name="Quote"/>
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  <w:LsdException Locked="false" Priority="30" QFormat="true"
 +
  Name="Intense Quote"/>
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  <w:LsdException Locked="false" Priority="66" Name="Medium List 2 Accent 1"/>
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  <w:LsdException Locked="false" Priority="67" Name="Medium Grid 1 Accent 1"/>
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  <w:LsdException Locked="false" Priority="68" Name="Medium Grid 2 Accent 1"/>
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  <w:LsdException Locked="false" Priority="69" Name="Medium Grid 3 Accent 1"/>
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  <w:LsdException Locked="false" Priority="70" Name="Dark List Accent 1"/>
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  <w:LsdException Locked="false" Priority="71" Name="Colorful Shading Accent 1"/>
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  <w:LsdException Locked="false" Priority="72" Name="Colorful List Accent 1"/>
 +
  <w:LsdException Locked="false" Priority="73" Name="Colorful Grid Accent 1"/>
 +
  <w:LsdException Locked="false" Priority="60" Name="Light Shading Accent 2"/>
 +
  <w:LsdException Locked="false" Priority="61" Name="Light List Accent 2"/>
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  <w:LsdException Locked="false" Priority="62" Name="Light Grid Accent 2"/>
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  <w:LsdException Locked="false" Priority="63" Name="Medium Shading 1 Accent 2"/>
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  <w:LsdException Locked="false" Priority="64" Name="Medium Shading 2 Accent 2"/>
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  <w:LsdException Locked="false" Priority="65" Name="Medium List 1 Accent 2"/>
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  <w:LsdException Locked="false" Priority="66" Name="Medium List 2 Accent 2"/>
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  <w:LsdException Locked="false" Priority="67" Name="Medium Grid 1 Accent 2"/>
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  <w:LsdException Locked="false" Priority="68" Name="Medium Grid 2 Accent 2"/>
 +
  <w:LsdException Locked="false" Priority="69" Name="Medium Grid 3 Accent 2"/>
 +
  <w:LsdException Locked="false" Priority="70" Name="Dark List Accent 2"/>
 +
  <w:LsdException Locked="false" Priority="71" Name="Colorful Shading Accent 2"/>
 +
  <w:LsdException Locked="false" Priority="72" Name="Colorful List Accent 2"/>
 +
  <w:LsdException Locked="false" Priority="73" Name="Colorful Grid Accent 2"/>
 +
  <w:LsdException Locked="false" Priority="60" Name="Light Shading Accent 3"/>
 +
  <w:LsdException Locked="false" Priority="61" Name="Light List Accent 3"/>
 +
  <w:LsdException Locked="false" Priority="62" Name="Light Grid Accent 3"/>
 +
  <w:LsdException Locked="false" Priority="63" Name="Medium Shading 1 Accent 3"/>
 +
  <w:LsdException Locked="false" Priority="64" Name="Medium Shading 2 Accent 3"/>
 +
  <w:LsdException Locked="false" Priority="65" Name="Medium List 1 Accent 3"/>
 +
  <w:LsdException Locked="false" Priority="66" Name="Medium List 2 Accent 3"/>
 +
  <w:LsdException Locked="false" Priority="67" Name="Medium Grid 1 Accent 3"/>
 +
  <w:LsdException Locked="false" Priority="68" Name="Medium Grid 2 Accent 3"/>
 +
  <w:LsdException Locked="false" Priority="69" Name="Medium Grid 3 Accent 3"/>
 +
  <w:LsdException Locked="false" Priority="70" Name="Dark List Accent 3"/>
 +
  <w:LsdException Locked="false" Priority="71" Name="Colorful Shading Accent 3"/>
 +
  <w:LsdException Locked="false" Priority="72" Name="Colorful List Accent 3"/>
 +
  <w:LsdException Locked="false" Priority="73" Name="Colorful Grid Accent 3"/>
 +
  <w:LsdException Locked="false" Priority="60" Name="Light Shading Accent 4"/>
 +
  <w:LsdException Locked="false" Priority="61" Name="Light List Accent 4"/>
 +
  <w:LsdException Locked="false" Priority="62" Name="Light Grid Accent 4"/>
 +
  <w:LsdException Locked="false" Priority="63" Name="Medium Shading 1 Accent 4"/>
 +
  <w:LsdException Locked="false" Priority="64" Name="Medium Shading 2 Accent 4"/>
 +
  <w:LsdException Locked="false" Priority="65" Name="Medium List 1 Accent 4"/>
 +
  <w:LsdException Locked="false" Priority="66" Name="Medium List 2 Accent 4"/>
 +
  <w:LsdException Locked="false" Priority="67" Name="Medium Grid 1 Accent 4"/>
 +
  <w:LsdException Locked="false" Priority="68" Name="Medium Grid 2 Accent 4"/>
 +
  <w:LsdException Locked="false" Priority="69" Name="Medium Grid 3 Accent 4"/>
 +
  <w:LsdException Locked="false" Priority="70" Name="Dark List Accent 4"/>
 +
  <w:LsdException Locked="false" Priority="71" Name="Colorful Shading Accent 4"/>
 +
  <w:LsdException Locked="false" Priority="72" Name="Colorful List Accent 4"/>
 +
  <w:LsdException Locked="false" Priority="73" Name="Colorful Grid Accent 4"/>
 +
  <w:LsdException Locked="false" Priority="60" Name="Light Shading Accent 5"/>
 +
  <w:LsdException Locked="false" Priority="61" Name="Light List Accent 5"/>
 +
  <w:LsdException Locked="false" Priority="62" Name="Light Grid Accent 5"/>
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  <w:LsdException Locked="false" Priority="63" Name="Medium Shading 1 Accent 5"/>
 +
  <w:LsdException Locked="false" Priority="64" Name="Medium Shading 2 Accent 5"/>
 +
  <w:LsdException Locked="false" Priority="65" Name="Medium List 1 Accent 5"/>
 +
  <w:LsdException Locked="false" Priority="66" Name="Medium List 2 Accent 5"/>
 +
  <w:LsdException Locked="false" Priority="67" Name="Medium Grid 1 Accent 5"/>
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  <w:LsdException Locked="false" Priority="68" Name="Medium Grid 2 Accent 5"/>
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  <w:LsdException Locked="false" Priority="69" Name="Medium Grid 3 Accent 5"/>
 +
  <w:LsdException Locked="false" Priority="70" Name="Dark List Accent 5"/>
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  <w:LsdException Locked="false" Priority="71" Name="Colorful Shading Accent 5"/>
 +
  <w:LsdException Locked="false" Priority="72" Name="Colorful List Accent 5"/>
 +
  <w:LsdException Locked="false" Priority="73" Name="Colorful Grid Accent 5"/>
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  <w:LsdException Locked="false" Priority="60" Name="Light Shading Accent 6"/>
 +
  <w:LsdException Locked="false" Priority="61" Name="Light List Accent 6"/>
 +
  <w:LsdException Locked="false" Priority="62" Name="Light Grid Accent 6"/>
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  <w:LsdException Locked="false" Priority="63" Name="Medium Shading 1 Accent 6"/>
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  <w:LsdException Locked="false" Priority="64" Name="Medium Shading 2 Accent 6"/>
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  <w:LsdException Locked="false" Priority="65" Name="Medium List 1 Accent 6"/>
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  <w:LsdException Locked="false" Priority="66" Name="Medium List 2 Accent 6"/>
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  <w:LsdException Locked="false" Priority="67" Name="Medium Grid 1 Accent 6"/>
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  <w:LsdException Locked="false" Priority="68" Name="Medium Grid 2 Accent 6"/>
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  <w:LsdException Locked="false" Priority="69" Name="Medium Grid 3 Accent 6"/>
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  <w:LsdException Locked="false" Priority="70" Name="Dark List Accent 6"/>
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  <w:LsdException Locked="false" Priority="71" Name="Colorful Shading Accent 6"/>
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  <w:LsdException Locked="false" Priority="72" Name="Colorful List Accent 6"/>
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  <w:LsdException Locked="false" Priority="73" Name="Colorful Grid Accent 6"/>
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  <w:LsdException Locked="false" Priority="19" QFormat="true"
 +
  Name="Subtle Emphasis"/>
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  <w:LsdException Locked="false" Priority="21" QFormat="true"
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  Name="Intense Emphasis"/>
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  <w:LsdException Locked="false" Priority="31" QFormat="true"
 +
  Name="Subtle Reference"/>
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  <w:LsdException Locked="false" Priority="32" QFormat="true"
 +
  Name="Intense Reference"/>
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  <w:LsdException Locked="false" Priority="33" QFormat="true" Name="Book Title"/>
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  <w:LsdException Locked="false" Priority="37" SemiHidden="true"
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  UnhideWhenUsed="true" Name="Bibliography"/>
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  <w:LsdException Locked="false" Priority="39" SemiHidden="true"
 +
  UnhideWhenUsed="true" QFormat="true" Name="TOC Heading"/>
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  <w:LsdException Locked="false" Priority="41" Name="Plain Table 1"/>
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  <w:LsdException Locked="false" Priority="42" Name="Plain Table 2"/>
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  <w:LsdException Locked="false" Priority="43" Name="Plain Table 3"/>
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  <w:LsdException Locked="false" Priority="44" Name="Plain Table 4"/>
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  <w:LsdException Locked="false" Priority="45" Name="Plain Table 5"/>
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  <w:LsdException Locked="false" Priority="40" Name="Grid Table Light"/>
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  <w:LsdException Locked="false" Priority="46" Name="Grid Table 1 Light"/>
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  <w:LsdException Locked="false" Priority="47" Name="Grid Table 2"/>
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  <w:LsdException Locked="false" Priority="48" Name="Grid Table 3"/>
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  <w:LsdException Locked="false" Priority="49" Name="Grid Table 4"/>
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  <w:LsdException Locked="false" Priority="50" Name="Grid Table 5 Dark"/>
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  <w:LsdException Locked="false" Priority="51" Name="Grid Table 6 Colorful"/>
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  <w:LsdException Locked="false" Priority="52" Name="Grid Table 7 Colorful"/>
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  <w:LsdException Locked="false" Priority="46"
 +
  Name="Grid Table 1 Light Accent 1"/>
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  <w:LsdException Locked="false" Priority="47" Name="Grid Table 2 Accent 1"/>
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  <w:LsdException Locked="false" Priority="48" Name="Grid Table 3 Accent 1"/>
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  <w:LsdException Locked="false" Priority="49" Name="Grid Table 4 Accent 1"/>
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  <w:LsdException Locked="false" Priority="50" Name="Grid Table 5 Dark Accent 1"/>
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  <w:LsdException Locked="false" Priority="51"
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  Name="Grid Table 6 Colorful Accent 1"/>
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  <w:LsdException Locked="false" Priority="52"
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  Name="Grid Table 7 Colorful Accent 1"/>
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  <w:LsdException Locked="false" Priority="46"
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  Name="Grid Table 1 Light Accent 2"/>
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  <w:LsdException Locked="false" Priority="47" Name="Grid Table 2 Accent 2"/>
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  <w:LsdException Locked="false" Priority="48" Name="Grid Table 3 Accent 2"/>
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  <w:LsdException Locked="false" Priority="49" Name="Grid Table 4 Accent 2"/>
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  <w:LsdException Locked="false" Priority="50" Name="Grid Table 5 Dark Accent 2"/>
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  <w:LsdException Locked="false" Priority="51"
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  Name="Grid Table 6 Colorful Accent 2"/>
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  <w:LsdException Locked="false" Priority="52"
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  Name="Grid Table 7 Colorful Accent 2"/>
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  <w:LsdException Locked="false" Priority="46"
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  Name="Grid Table 1 Light Accent 3"/>
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  <w:LsdException Locked="false" Priority="47" Name="Grid Table 2 Accent 3"/>
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  <w:LsdException Locked="false" Priority="48" Name="Grid Table 3 Accent 3"/>
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  <w:LsdException Locked="false" Priority="49" Name="Grid Table 4 Accent 3"/>
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  <w:LsdException Locked="false" Priority="50" Name="Grid Table 5 Dark Accent 3"/>
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  <w:LsdException Locked="false" Priority="51"
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  Name="Grid Table 6 Colorful Accent 3"/>
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  <w:LsdException Locked="false" Priority="52"
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  Name="Grid Table 7 Colorful Accent 3"/>
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  <w:LsdException Locked="false" Priority="46"
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  Name="Grid Table 1 Light Accent 4"/>
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  <w:LsdException Locked="false" Priority="47" Name="Grid Table 2 Accent 4"/>
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  <w:LsdException Locked="false" Priority="48" Name="Grid Table 3 Accent 4"/>
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  <w:LsdException Locked="false" Priority="49" Name="Grid Table 4 Accent 4"/>
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  <w:LsdException Locked="false" Priority="50" Name="Grid Table 5 Dark Accent 4"/>
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  <w:LsdException Locked="false" Priority="51"
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  Name="Grid Table 6 Colorful Accent 4"/>
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  <w:LsdException Locked="false" Priority="52"
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  Name="Grid Table 7 Colorful Accent 4"/>
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  <w:LsdException Locked="false" Priority="46"
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  Name="Grid Table 1 Light Accent 5"/>
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  <w:LsdException Locked="false" Priority="48" Name="Grid Table 3 Accent 5"/>
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  <w:LsdException Locked="false" Priority="49" Name="Grid Table 4 Accent 5"/>
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  <w:LsdException Locked="false" Priority="50" Name="Grid Table 5 Dark Accent 5"/>
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  <w:LsdException Locked="false" Priority="51"
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  Name="Grid Table 6 Colorful Accent 5"/>
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  <w:LsdException Locked="false" Priority="52"
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  Name="Grid Table 7 Colorful Accent 5"/>
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  <w:LsdException Locked="false" Priority="46"
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  Name="Grid Table 1 Light Accent 6"/>
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  <w:LsdException Locked="false" Priority="47" Name="Grid Table 2 Accent 6"/>
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  <w:LsdException Locked="false" Priority="48" Name="Grid Table 3 Accent 6"/>
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  <w:LsdException Locked="false" Priority="49" Name="Grid Table 4 Accent 6"/>
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  <w:LsdException Locked="false" Priority="50" Name="Grid Table 5 Dark Accent 6"/>
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  <w:LsdException Locked="false" Priority="51"
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  Name="Grid Table 6 Colorful Accent 6"/>
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  <w:LsdException Locked="false" Priority="52"
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  Name="Grid Table 7 Colorful Accent 6"/>
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  <w:LsdException Locked="false" Priority="46" Name="List Table 1 Light"/>
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  <w:LsdException Locked="false" Priority="47" Name="List Table 2"/>
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  <w:LsdException Locked="false" Priority="48" Name="List Table 3"/>
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  <w:LsdException Locked="false" Priority="49" Name="List Table 4"/>
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  <w:LsdException Locked="false" Priority="50" Name="List Table 5 Dark"/>
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  <w:LsdException Locked="false" Priority="51" Name="List Table 6 Colorful"/>
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  <w:LsdException Locked="false" Priority="52" Name="List Table 7 Colorful"/>
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  <w:LsdException Locked="false" Priority="46"
 +
  Name="List Table 1 Light Accent 1"/>
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  <w:LsdException Locked="false" Priority="47" Name="List Table 2 Accent 1"/>
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  <w:LsdException Locked="false" Priority="48" Name="List Table 3 Accent 1"/>
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  <w:LsdException Locked="false" Priority="49" Name="List Table 4 Accent 1"/>
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  <w:LsdException Locked="false" Priority="50" Name="List Table 5 Dark Accent 1"/>
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  <w:LsdException Locked="false" Priority="51"
 +
  Name="List Table 6 Colorful Accent 1"/>
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  <w:LsdException Locked="false" Priority="52"
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  Name="List Table 7 Colorful Accent 1"/>
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  <w:LsdException Locked="false" Priority="46"
 +
  Name="List Table 1 Light Accent 2"/>
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  <w:LsdException Locked="false" Priority="47" Name="List Table 2 Accent 2"/>
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  <w:LsdException Locked="false" Priority="48" Name="List Table 3 Accent 2"/>
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  <w:LsdException Locked="false" Priority="49" Name="List Table 4 Accent 2"/>
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  <w:LsdException Locked="false" Priority="50" Name="List Table 5 Dark Accent 2"/>
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  <w:LsdException Locked="false" Priority="51"
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  Name="List Table 6 Colorful Accent 2"/>
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  <w:LsdException Locked="false" Priority="52"
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  Name="List Table 7 Colorful Accent 2"/>
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  <w:LsdException Locked="false" Priority="46"
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  Name="List Table 1 Light Accent 3"/>
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  <w:LsdException Locked="false" Priority="47" Name="List Table 2 Accent 3"/>
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  <w:LsdException Locked="false" Priority="48" Name="List Table 3 Accent 3"/>
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  <w:LsdException Locked="false" Priority="49" Name="List Table 4 Accent 3"/>
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  <w:LsdException Locked="false" Priority="50" Name="List Table 5 Dark Accent 3"/>
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  <w:LsdException Locked="false" Priority="51"
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  Name="List Table 6 Colorful Accent 3"/>
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  <w:LsdException Locked="false" Priority="52"
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  Name="List Table 7 Colorful Accent 3"/>
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  <w:LsdException Locked="false" Priority="46"
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  Name="List Table 1 Light Accent 4"/>
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  <w:LsdException Locked="false" Priority="47" Name="List Table 2 Accent 4"/>
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  <w:LsdException Locked="false" Priority="48" Name="List Table 3 Accent 4"/>
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  <w:LsdException Locked="false" Priority="49" Name="List Table 4 Accent 4"/>
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  <w:LsdException Locked="false" Priority="50" Name="List Table 5 Dark Accent 4"/>
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  <w:LsdException Locked="false" Priority="51"
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  Name="List Table 6 Colorful Accent 4"/>
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  <w:LsdException Locked="false" Priority="52"
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  Name="List Table 7 Colorful Accent 4"/>
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  <w:LsdException Locked="false" Priority="46"
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  Name="List Table 1 Light Accent 5"/>
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  <w:LsdException Locked="false" Priority="47" Name="List Table 2 Accent 5"/>
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  <w:LsdException Locked="false" Priority="48" Name="List Table 3 Accent 5"/>
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  <w:LsdException Locked="false" Priority="49" Name="List Table 4 Accent 5"/>
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  <w:LsdException Locked="false" Priority="50" Name="List Table 5 Dark Accent 5"/>
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  <w:LsdException Locked="false" Priority="51"
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  Name="List Table 6 Colorful Accent 5"/>
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  <w:LsdException Locked="false" Priority="52"
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  Name="List Table 7 Colorful Accent 5"/>
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  <w:LsdException Locked="false" Priority="46"
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  Name="List Table 1 Light Accent 6"/>
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  <w:LsdException Locked="false" Priority="47" Name="List Table 2 Accent 6"/>
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  <w:LsdException Locked="false" Priority="48" Name="List Table 3 Accent 6"/>
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  <w:LsdException Locked="false" Priority="49" Name="List Table 4 Accent 6"/>
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  <w:LsdException Locked="false" Priority="50" Name="List Table 5 Dark Accent 6"/>
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  <w:LsdException Locked="false" Priority="51"
 +
  Name="List Table 6 Colorful Accent 6"/>
 +
  <w:LsdException Locked="false" Priority="52"
 +
  Name="List Table 7 Colorful Accent 6"/>
 +
</w:LatentStyles>
 +
</xml><![endif]-->
 +
<style>
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/* Font Definitions */
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@font-face
 +
{font-family:"Cambria Math";
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panose-1:2 4 5 3 5 4 6 3 2 4;
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mso-font-charset:1;
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mso-generic-font-family:roman;
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mso-font-format:other;
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mso-font-pitch:variable;
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mso-font-signature:0 0 0 0 0 0;}
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@font-face
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{font-family:Calibri;
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panose-1:2 15 5 2 2 2 4 3 2 4;
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mso-font-charset:162;
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mso-generic-font-family:swiss;
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mso-font-pitch:variable;
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mso-font-signature:-536870145 1073786111 1 0 415 0;}
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@font-face
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{font-family:Tahoma;
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panose-1:2 11 6 4 3 5 4 4 2 4;
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mso-font-charset:162;
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mso-generic-font-family:swiss;
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mso-font-pitch:variable;
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mso-font-signature:-520081665 -1073717157 41 0 66047 0;}
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/* Style Definitions */
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p.MsoNormal, li.MsoNormal, div.MsoNormal
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{mso-style-unhide:no;
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mso-style-qformat:yes;
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mso-style-parent:"";
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margin-top:0cm;
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margin-right:0cm;
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margin-bottom:10.0pt;
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margin-left:0cm;
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line-height:115%;
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mso-pagination:widow-orphan;
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font-size:11.0pt;
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font-family:"Calibri","sans-serif";
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mso-fareast-font-family:"Times New Roman";
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mso-fareast-theme-font:minor-fareast;
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mso-bidi-font-family:"Times New Roman";}
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p.MsoAcetate, li.MsoAcetate, div.MsoAcetate
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{mso-style-noshow:yes;
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mso-style-priority:99;
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mso-style-link:"Balon Metni Char";
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margin:0cm;
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margin-bottom:.0001pt;
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mso-pagination:widow-orphan;
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mso-fareast-font-family:"Times New Roman";
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mso-fareast-theme-font:minor-fareast;}
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p.MsoListParagraph, li.MsoListParagraph, div.MsoListParagraph
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mso-style-qformat:yes;
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mso-pagination:widow-orphan;
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mso-fareast-font-family:"Times New Roman";
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mso-fareast-theme-font:minor-fareast;
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mso-bidi-font-family:"Times New Roman";}
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span.BalonMetniChar
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{mso-style-name:"Balon Metni Char";
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mso-style-noshow:yes;
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mso-style-priority:99;
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mso-style-unhide:no;
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mso-style-link:"Balon Metni";
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mso-hansi-font-family:Tahoma;
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mso-bidi-font-family:Tahoma;}
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p.msolistparagraphcxspfirst, li.msolistparagraphcxspfirst, div.msolistparagraphcxspfirst
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 +
 
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<div class=WordSection1>
 +
 
 +
<p class=MsoNormal><b><u><span lang=EN-US style='color:red;mso-ansi-language:
 +
EN-US'>Spectrophotometric Absorption Measurements: </span></u></b><span
 +
lang=EN-US style='mso-ansi-language:EN-US'>&nbsp;We used </span><b>Thermo Sc</b><b><span
 +
lang=EN-US style='mso-ansi-language:EN-US'>ientific Varioskan Flash Multimode
 +
Reader for our absorption measurements.</span></b></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='mso-ansi-language:EN-US'>1)
 +
Measurements of Dyes:</span></p>
 +
 
 +
<p class=MsoNormal><span style='mso-no-proof:yes'><img width=604 height=440
 +
id="_x0000_i1031" src="https://static.igem.org/mediawiki/2014hs/8/88/Fimage001.jpg"
 +
alt="https://static.igem.org/mediawiki/2014hs/8/88/Fimage001.jpg"></span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='mso-ansi-language:EN-US'>Graphic 1:
 +
This graphic shows our absorption rate of our dye (Methylene Blue) in different
 +
concentrations. ( 1X=8 uM, 10X=80uM, 100X=800uM, 1000X=uM) </span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='mso-ansi-language:EN-US'>In the
 +
Graphic 1, we have got the correct absorption rates for every concentration of
 +
Methylene Blue dyes that correlates with the same results in the literature.</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='mso-ansi-language:EN-US'>2)
 +
Preparations of Lysates : <o:p></o:p></span></p>
 +
 
 +
<p class=MsoNormal><span style='font-size:11.5pt;line-height:115%;font-family:
 +
"Arial","sans-serif";color:black'>The verified plasmids were then cultivated in
 +
liquid culture for isolation. The 50 mL’s of bacteria containing the parts
 +
BBa_K1291071 and BBa_K1291299 were centrifuged at 4100 rpm for 5 minutes, the
 +
supernatant was discarded and then lysis solution was added. After 10 minutes
 +
of incubation in ice, the lysates were sonicated in the minimum activity level.
 +
After a spin of 15000 rpm for 10 minutes, the supernatant was transferred to
 +
eppendorphs and dye was added; the solution was boiled at 95</span></span><span
 +
style='font-size:11.5pt;line-height:115%;font-family:"Arial","sans-serif";
 +
color:#545454; '><span style='orphans: auto;text-align:start;
 +
widows: auto;-webkit-text-stroke-width: 0px;white-space:pre-wrap;word-spacing:
 +
0px'> °C for 5 minutes. The lysate were stored at -20 °C.</span></span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='mso-ansi-language:EN-US'>3)
 +
Decolorization Assay:</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='mso-ansi-language:EN-US'>In this
 +
assay we mixed our liquid cell culture and also our lysates with dyes. All the
 +
concentrations of the solutions are adjusted according to the literature. We
 +
measured the first absorptions of the samples and compared it with the 2<sup>nd</sup>
 +
measurement in the end of 1<sup>st</sup> hour.</span></p>
 +
 
 +
<p class=MsoNormal><span style='mso-no-proof:yes'><img width=604 height=300
 +
id="_x0000_i1030" src="https://static.igem.org/mediawiki/2014hs/f/fb/Fimage002.jpg"
 +
alt="https://static.igem.org/mediawiki/2014hs/f/fb/Fimage002.jpg"></span>&nbsp;</p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='mso-ansi-language:EN-US'>Graphic 2:
 +
This graphic shows our absorption rate of our mixed sample that includes
 +
Lysates of our BL-21 Bacteria that have LiP gene and&nbsp; Methylene Blue
 +
(80uM) dyes and for the activate the LiP we also added Hydrogen Peroxide
 +
(800uM) measured in the 1 hour.</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='mso-ansi-language:EN-US'>The result
 +
of Graphic-2 is that our Lysates of BL-21 Bacteria that have LiP gene degraded
 +
the Methylene Blue (80uM) dye with approximately %83,3 efficiency.</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='mso-ansi-language:EN-US'>&nbsp;</span></p>
 +
 
 +
<p class=MsoNormal><span style='mso-no-proof:yes'><img width=605 height=516
 +
id="_x0000_i1029" src="https://static.igem.org/mediawiki/2014hs/7/7d/Fimage003.jpg"
 +
alt="https://static.igem.org/mediawiki/2014hs/7/7d/Fimage003.jpg"></span><span
 +
lang=EN-US style='mso-ansi-language:EN-US'>Graphic 3: This graphic shows our
 +
absorption rate of our mixed samples that includes Lysates of our BL-21
 +
Bacteria that have HRP gene and other Bacteria that have LiP gene and Methyl
 +
Green (150uM) dyes and for the activate the HRP and LiP we also added Hydrogen
 +
Peroxide (8.8 uM-HRP), (0,176 uM-LiP) concentrations that measured in the
 +
different time periods. </span></p>
 +
 
 +
<p class=MsoNormal><u><span lang=EN-US style='color:red;mso-ansi-language:EN-US'>The
 +
result of Graphic-3 is that our Lysates of BL-21 Bacteria that have HRP gene
 +
and other Bacteria that have LiP gene degraded the Methyl Green (8.8 uM) dye
 +
with approximately %71,4 efficiency in the end of the 7 hours.</span></u></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='mso-ansi-language:EN-US'>&nbsp;</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='mso-ansi-language:EN-US'>&nbsp;</span></p>
 +
 
 +
<p class=MsoNormal><span style='mso-no-proof:yes'><img width=475 height=304
 +
id="_x0000_i1028" src="https://static.igem.org/mediawiki/2014hs/e/ed/Fimage004.png"
 +
alt="https://static.igem.org/mediawiki/2014hs/e/ed/Fimage004.png"></span><span
 +
lang=EN-US style='mso-ansi-language:EN-US'>.</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='mso-ansi-language:EN-US'>Graphic 4:
 +
This graphic shows our absorption rate of our mixed samples that includes
 +
Lysates of our BL-21 Bacteria that have HRP gene and other Bacteria that have
 +
LiP gene and Methyl Green (150uM) dyes and for the activate the HRP and LiP we
 +
also added Hydrogen Peroxide (8.8 uM-HRP), (0,176 uM-LiP) concentrations that
 +
measured in the beginning of the experiment and in the 1<sup>st</sup> hour. </span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='mso-ansi-language:EN-US'>The result
 +
of Graphic-4 is that our Lysates of BL-21 Bacteria that have HRP gene and other
 +
Bacteria that have LiP gene degraded the Methyl Green (8.8 uM) dye with
 +
approximately %71,4 efficiency. <u><span style='color:red'>Also in this graphic
 +
the most important point that our control group BL-21 bacteria that don’t
 +
include our genes of enzymes(HRP or LiP) couldn’t degrade the Methyl Green (8.8
 +
uM) dye.</span></u></span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='mso-ansi-language:EN-US'>&nbsp;</span></p>
 +
 
 +
<p class=MsoNormal><span style='mso-no-proof:yes'><img width=610 height=320
 +
id="_x0000_i1027" src="https://static.igem.org/mediawiki/2014hs/a/a9/Fimage005.jpg"
 +
alt="https://static.igem.org/mediawiki/2014hs/a/a9/Fimage005.jpg"></span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='mso-ansi-language:EN-US'>Graphic 5:
 +
This graphic shows our absorption rate of our mixed sample that includes liquid
 +
culture &nbsp;of our BL-21 Bacteria that have LiP gene and&nbsp; Methylene Blue
 +
(80uM) dyes and for the activate the LiP we also added Hydrogen Peroxide
 +
(800uM) measured in the 1 hour.</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='mso-ansi-language:EN-US'>The result
 +
of Graphic-5 is that our Liquid cultures of BL-21 Bacteria that have LiP gene
 +
also degraded the Methylene Blue (80uM) dye with approximately %70 efficiency. </span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='mso-ansi-language:EN-US'>&nbsp;</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='mso-ansi-language:EN-US'>&nbsp;</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='mso-ansi-language:EN-US'>&nbsp;</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='mso-ansi-language:EN-US'>&nbsp;</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='mso-ansi-language:EN-US'>&nbsp;</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='mso-ansi-language:EN-US'>&nbsp;</span></p>
 +
 
 +
<p class=MsoNormal><span style='mso-no-proof:yes'><img width=603 height=359
 +
id="_x0000_i1026" src="https://static.igem.org/mediawiki/2014hs/e/eb/Fimage006.jpg"
 +
alt="https://static.igem.org/mediawiki/2014hs/e/eb/Fimage006.jpg"></span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='mso-ansi-language:EN-US'>Graphic 6:
 +
This graphic shows our absorption rate of our mixed sample that includes liquid
 +
culture&nbsp; of our BL-21 Bacteria that have HRP gene and&nbsp; Methylene Blue
 +
(80uM) dyes and for the activate the HRP we also added Hydrogen Peroxide
 +
(800uM) measured in the 1 hour.</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='mso-ansi-language:EN-US'>The result
 +
of Graphic-6 is that our Liquid cultures of BL-21 Bacteria that have LiP gene
 +
also degraded the Methylene Blue (80uM) dye with approximately %65 efficiency.</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='mso-ansi-language:EN-US'>&nbsp;</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='mso-ansi-language:EN-US'>&nbsp;</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='mso-ansi-language:EN-US'>The Results
 +
of Graphic 5 and 6 show that our liquid culture of our BL-21 Bacteria that have
 +
HRP gene and other Bacteria that have LiP gene could degraded the Methylene
 +
Blue (80uM) dye but our Lysates of our BL-21 Bacteria that have HRP gene and
 +
other Bacteria that have LiP gene degraded the Methylene Blue (80uM) dye more
 +
efficiently. This results gave us an opinion that when our enzymes have further
 +
interaction with dyes they could degrade much more.</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='mso-ansi-language:EN-US'>&nbsp;</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='mso-ansi-language:EN-US'>&nbsp;</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='mso-ansi-language:EN-US'>&nbsp;</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='mso-ansi-language:EN-US'>&nbsp;</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='mso-ansi-language:EN-US'>&nbsp;</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='mso-ansi-language:EN-US'>&nbsp;</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='mso-ansi-language:EN-US'>&nbsp;</span></p>
 +
 
 +
<p class=MsoNormal><span style='mso-no-proof:yes'><img width=604 height=283
 +
id="_x0000_i1025" src="https://static.igem.org/mediawiki/2014hs/6/60/Fimage007.jpg"
 +
alt="https://static.igem.org/mediawiki/2014hs/6/60/Fimage007.jpg"></span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='mso-ansi-language:EN-US'>&nbsp;</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='mso-ansi-language:EN-US'>Graphic 7:
 +
This graphic shows our absorption rate of our mixed sample that includes liquid
 +
culture&nbsp; of our Control BL-21 bacteria that don’t have any HRP or LiP gene
 +
and&nbsp; Methylene Blue (80uM) dyes and for the activate the any peroxidases,
 +
Control BL-21 bacteria have, we also added Hydrogen Peroxide (800uM) measured
 +
in the 1 hour.</span></p>
 +
 
 +
<p class=MsoNormal><span lang=EN-US style='mso-ansi-language:EN-US'>The result
 +
of Graphic-7 is that our Liquid culture of BL-21 Bacteria that don’t have any
 +
HRP or LiP gene could degrade the Methylene Blue (80uM) dye with approximately
 +
% 18 efficiency. This result show that Control bacteria can degrade the the
 +
Methylene Blue (80uM) dye but at the very low percentage when compared with our
 +
liquid culture BL-21 Bacteria that have HRP gene and other Bacteria that have
 +
LiP gene so we thought that some peroxidases also can degrade the dyes but our
 +
enzymes HRP and LiP have very high catalytic ability for the our
 +
dyes(Methlylene Blue and Methyl Green). These results and graphics show that
 +
our enzymes are functional and working properly.</span></p>
 +
 
 +
</div>
 +
 
 +
</body>
 +
 
 +
 
</head>
</head>
</html>
</html>

Latest revision as of 17:35, 26 April 2015

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Spectrophotometric Absorption Measurements:  We used Thermo Scientific Varioskan Flash Multimode Reader for our absorption measurements.

1) Measurements of Dyes:

https://static.igem.org/mediawiki/2014hs/8/88/Fimage001.jpg

Graphic 1: This graphic shows our absorption rate of our dye (Methylene Blue) in different concentrations. ( 1X=8 uM, 10X=80uM, 100X=800uM, 1000X=uM)

In the Graphic 1, we have got the correct absorption rates for every concentration of Methylene Blue dyes that correlates with the same results in the literature.

2) Preparations of Lysates :

The verified plasmids were then cultivated in liquid culture for isolation. The 50 mL’s of bacteria containing the parts BBa_K1291071 and BBa_K1291299 were centrifuged at 4100 rpm for 5 minutes, the supernatant was discarded and then lysis solution was added. After 10 minutes of incubation in ice, the lysates were sonicated in the minimum activity level. After a spin of 15000 rpm for 10 minutes, the supernatant was transferred to eppendorphs and dye was added; the solution was boiled at 95 °C for 5 minutes. The lysate were stored at -20 °C.

3) Decolorization Assay:

In this assay we mixed our liquid cell culture and also our lysates with dyes. All the concentrations of the solutions are adjusted according to the literature. We measured the first absorptions of the samples and compared it with the 2nd measurement in the end of 1st hour.

https://static.igem.org/mediawiki/2014hs/f/fb/Fimage002.jpg 

Graphic 2: This graphic shows our absorption rate of our mixed sample that includes Lysates of our BL-21 Bacteria that have LiP gene and  Methylene Blue (80uM) dyes and for the activate the LiP we also added Hydrogen Peroxide (800uM) measured in the 1 hour.

The result of Graphic-2 is that our Lysates of BL-21 Bacteria that have LiP gene degraded the Methylene Blue (80uM) dye with approximately %83,3 efficiency.

 

https://static.igem.org/mediawiki/2014hs/7/7d/Fimage003.jpgGraphic 3: This graphic shows our absorption rate of our mixed samples that includes Lysates of our BL-21 Bacteria that have HRP gene and other Bacteria that have LiP gene and Methyl Green (150uM) dyes and for the activate the HRP and LiP we also added Hydrogen Peroxide (8.8 uM-HRP), (0,176 uM-LiP) concentrations that measured in the different time periods.

The result of Graphic-3 is that our Lysates of BL-21 Bacteria that have HRP gene and other Bacteria that have LiP gene degraded the Methyl Green (8.8 uM) dye with approximately %71,4 efficiency in the end of the 7 hours.

 

 

https://static.igem.org/mediawiki/2014hs/e/ed/Fimage004.png.

Graphic 4: This graphic shows our absorption rate of our mixed samples that includes Lysates of our BL-21 Bacteria that have HRP gene and other Bacteria that have LiP gene and Methyl Green (150uM) dyes and for the activate the HRP and LiP we also added Hydrogen Peroxide (8.8 uM-HRP), (0,176 uM-LiP) concentrations that measured in the beginning of the experiment and in the 1st hour.

The result of Graphic-4 is that our Lysates of BL-21 Bacteria that have HRP gene and other Bacteria that have LiP gene degraded the Methyl Green (8.8 uM) dye with approximately %71,4 efficiency. Also in this graphic the most important point that our control group BL-21 bacteria that don’t include our genes of enzymes(HRP or LiP) couldn’t degrade the Methyl Green (8.8 uM) dye.

 

https://static.igem.org/mediawiki/2014hs/a/a9/Fimage005.jpg

Graphic 5: This graphic shows our absorption rate of our mixed sample that includes liquid culture  of our BL-21 Bacteria that have LiP gene and  Methylene Blue (80uM) dyes and for the activate the LiP we also added Hydrogen Peroxide (800uM) measured in the 1 hour.

The result of Graphic-5 is that our Liquid cultures of BL-21 Bacteria that have LiP gene also degraded the Methylene Blue (80uM) dye with approximately %70 efficiency.

 

 

 

 

 

 

https://static.igem.org/mediawiki/2014hs/e/eb/Fimage006.jpg

Graphic 6: This graphic shows our absorption rate of our mixed sample that includes liquid culture  of our BL-21 Bacteria that have HRP gene and  Methylene Blue (80uM) dyes and for the activate the HRP we also added Hydrogen Peroxide (800uM) measured in the 1 hour.

The result of Graphic-6 is that our Liquid cultures of BL-21 Bacteria that have LiP gene also degraded the Methylene Blue (80uM) dye with approximately %65 efficiency.

 

 

The Results of Graphic 5 and 6 show that our liquid culture of our BL-21 Bacteria that have HRP gene and other Bacteria that have LiP gene could degraded the Methylene Blue (80uM) dye but our Lysates of our BL-21 Bacteria that have HRP gene and other Bacteria that have LiP gene degraded the Methylene Blue (80uM) dye more efficiently. This results gave us an opinion that when our enzymes have further interaction with dyes they could degrade much more.

 

 

 

 

 

 

 

https://static.igem.org/mediawiki/2014hs/6/60/Fimage007.jpg

 

Graphic 7: This graphic shows our absorption rate of our mixed sample that includes liquid culture  of our Control BL-21 bacteria that don’t have any HRP or LiP gene and  Methylene Blue (80uM) dyes and for the activate the any peroxidases, Control BL-21 bacteria have, we also added Hydrogen Peroxide (800uM) measured in the 1 hour.

The result of Graphic-7 is that our Liquid culture of BL-21 Bacteria that don’t have any HRP or LiP gene could degrade the Methylene Blue (80uM) dye with approximately % 18 efficiency. This result show that Control bacteria can degrade the the Methylene Blue (80uM) dye but at the very low percentage when compared with our liquid culture BL-21 Bacteria that have HRP gene and other Bacteria that have LiP gene so we thought that some peroxidases also can degrade the dyes but our enzymes HRP and LiP have very high catalytic ability for the our dyes(Methlylene Blue and Methyl Green). These results and graphics show that our enzymes are functional and working properly.

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