Difference between revisions of "Team:Freiburg/Protocols/MesurementGFP"

(Created page with "{{Freiburg/CSS}} {{Freiburg/Menubar}} {{Freiburg/LabjournalCSS}} {{Freiburg/wiki_content_start}} <html><style> →‎========= BEGIN: style for navigation bar ==========: #noteb...")
 
Line 39: Line 39:
 
<hr/>
 
<hr/>
 
</div>
 
</div>
−
<h4>Terrific Broth</h4>
+
 
 +
 
 +
 
 +
 
 +
<h4>Measurement of expressed GFP</h4>
 
<div class="level4">
 
<div class="level4">
−
<div class="table sectionedit2"><table class="inline">
+
<ul>
−
<tr class="row0">
+
<li class="level1"><div class="li">All fluorescence measurements were performed with a microplate reader in blach plates with transparent bottom </div>
−
<th class="col0">Component </th><th class="col1"> for 1 litre medium </th><td class="col2"> <b>remarks</b> </td>
+
−
</tr>
+
−
<tr class="row1">
+
−
<td class="col0"> Monopotassium phosphate </td><td class="col1"> 5.6 g </td><td class="col2">  </td>
+
−
<tr class="row2">
+
−
<td class="col0"> Potassium phosphate dibasic  </td><td class="col1"> 28.9 g </td><td class="col2">  </td>
+
−
<tr class="row3">
+
−
<td class="col0"> yeast extract </td><td class="col1"> 10  </td>
+
−
</tr>
+
−
<tr class="row4">
+
−
<td class="col0"> Thiamine </td><td class="col1"> 10 mg </td><td class="col2"> added after sterilisation (filter sterilized) </td>
+
−
</tr>
+
−
<tr class="row5">
+
−
<td class="col0"> Glucose (20%) </td><td class="col1"> 40ml </td><td class="col2"> added after sterilisation (filter sterilized) </td>
+
−
</tr>
+
−
</table></div>
+
−
<!-- EDIT2 TABLE [412-920] -->
+
−
</div>
+
−
<li class="level1"><div class="li"> autoclave medium than add filter sterilized Thiamine and Glucose</div>
+
 
</li>
 
</li>
−
<li class="level1"><div class="li"> Store at 4°C</div>
+
<li class="level1"><div class="li">As Exitation wavelength 480nm were choosen </div>
−
<ol>
+
</li>
 +
<li class="level1"><div class="li">Emission was measured at 520nm or a full spectrum was taken from 500 to 700 nm</div>
 +
</li>
 +
<li class="level1"><div class="li">Reactions were either performed in a tube and transferred to the microplate after the cell-free reaction or directly performed inside the platereader </div>
 +
</li>
 +
<li class="level1"><div class="li">when the reaction was performed inside the Platreader the wells were covered with foil to avoid evaporation</div>
 +
</li>
 +
<li class="level1"><div class="li"> After the reaction and Measurement samples were stored at 4°C for further folding or directly frozen at -20°C</div>
 +
</ul>
 +
 
  
 
<div class="tags"><span>
 
<div class="tags"><span>

Revision as of 22:56, 14 September 2015

""

<< Back

Measurement of GFP Expression

material: chemicals, used kits, …
duration: 90 min

Measurement of expressed GFP

  • All fluorescence measurements were performed with a microplate reader in blach plates with transparent bottom
  • As Exitation wavelength 480nm were choosen
  • Emission was measured at 520nm or a full spectrum was taken from 500 to 700 nm
  • Reactions were either performed in a tube and transferred to the microplate after the cell-free reaction or directly performed inside the platereader
  • when the reaction was performed inside the Platreader the wells were covered with foil to avoid evaporation
  • After the reaction and Measurement samples were stored at 4°C for further folding or directly frozen at -20°C
,