Difference between revisions of "Team:Michigan/Notebook/September"

 
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<h2 style="text-align: center">Septebmer</h2>
+
<h2 style="text-align: center">September</h2>
 +
<p>
 +
Work plan:
 +
<br><br>
 +
LacZ into Thrombin plasmid:
 +
<br>
 +
<ul>
 +
  <li>cloning not working probably because the insert is almost as big as backbone let’s transform LacZ again</li>
 +
  <li> transform in vitro LacZ part again</li>
 +
  <li>plasmid is ready, digest with HindiIII and A</li>
 +
  <li>gel extract plasmid, measure dna concentration</li>
 +
</ul>
  
 +
<br>
 +
LacZ into Gliadin plasmid:
 +
<br>
 +
Pfldh plasmid with GFP:
 +
<br>measure fluorescence
 +
<br><br>
 +
Things to order:
 +
<br>
 +
<ul>
 +
  <li>aptamer and trigger for Gliadin</li>
 +
  <li> switch for Gliadin with GFP (check design folder)</li>
 +
  <li>PfLDH protein</li>
 +
  <li>aptamer and trigger for Pfldh</li>
 +
  <li>aptamer and trigger for Kanamycin</li>
 +
</ul>
 +
 +
09/07/2015
 +
<br><br>
 +
we wanted to replicate results from previous experiment and try to detect lower thrombin concentrations to prove the sensitivity of our system
 +
<br><br>
 +
 +
1 = Negative control (switch 1.0 + trigger-aptamer No Thrombin)<br>
 +
2 = Positive control (switch 1.0 + trigger)<br>
 +
3= Switch 1.0 + trigger + aptamer + 25uM Thrombin<br>
 +
4= Switch 1.0 + trigger + aptamer + 5uM Thrombin<br>
 +
5= Switch 1.0 + trigger + aptamer + 2.5uM Thrombin<br>
 +
6= Switch 1.0 + trigger + aptamer + 1.0uM Thrombin<br>
 +
7= Switch 1.0 + trigger + aptamer + 0.5uM Thrombin<br>
 +
8= Switch 1.0 + trigger + aptamer + 0.1uM Thrombin<br>
 +
<br><br><br>
 +
 +
<img src="https://static.igem.org/mediawiki/2015/3/36/SwitchThrombin.png" alt="MostImportant" style="width:504px;height:428px;">
 +
<br><br>
 +
<img src="https://static.igem.org/mediawiki/2015/1/19/Gliadin_titration.png" alt="Gliadin" style="width:504px;height:428px;">
 +
<br><br>
 +
<img src="https://static.igem.org/mediawiki/2015/a/a7/Table_thing.png" alt="TableThing" style="width:504px;height:428px;">
 +
<br><br>
  
 
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Latest revision as of 03:58, 19 September 2015

September

Work plan:

LacZ into Thrombin plasmid:

  • cloning not working probably because the insert is almost as big as backbone let’s transform LacZ again
  • transform in vitro LacZ part again
  • plasmid is ready, digest with HindiIII and A
  • gel extract plasmid, measure dna concentration

LacZ into Gliadin plasmid:
Pfldh plasmid with GFP:
measure fluorescence

Things to order:
  • aptamer and trigger for Gliadin
  • switch for Gliadin with GFP (check design folder)
  • PfLDH protein
  • aptamer and trigger for Pfldh
  • aptamer and trigger for Kanamycin
09/07/2015

we wanted to replicate results from previous experiment and try to detect lower thrombin concentrations to prove the sensitivity of our system

1 = Negative control (switch 1.0 + trigger-aptamer No Thrombin)
2 = Positive control (switch 1.0 + trigger)
3= Switch 1.0 + trigger + aptamer + 25uM Thrombin
4= Switch 1.0 + trigger + aptamer + 5uM Thrombin
5= Switch 1.0 + trigger + aptamer + 2.5uM Thrombin
6= Switch 1.0 + trigger + aptamer + 1.0uM Thrombin
7= Switch 1.0 + trigger + aptamer + 0.5uM Thrombin
8= Switch 1.0 + trigger + aptamer + 0.1uM Thrombin



MostImportant

Gliadin

TableThing