Difference between revisions of "Team:Munich/cas13a.html"

(Created page with "<div class="event-info"> <h3>4/24</h3> <h4>restriction digest p2ct_his_MBP_LBU_c2c2_WT</h4> <table class="table table-borderless"> <tr> <td>Participants:</td>...")
 
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<html>
 
<div class="event-info">
 
<div class="event-info">
  
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<h4>Assembling p2ct_his_MBP_LBU_c2c2_gRNA-EndoA</h4>
−
<h3>4/24</h3>
+
<em>2018/04/24 - 2018/04/26</em>
−
<h4>restriction digest p2ct_his_MBP_LBU_c2c2_WT</h4>
+
 
<table class="table table-borderless">
 
<table class="table table-borderless">
 
     <tr>
 
     <tr>
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     <tr>
 
     <tr>
 
       <td>Protocol:</td>
 
       <td>Protocol:</td>
−
       <td>restriction digest</td>
+
       <td>restriction digest, gibson ligation, chem trafo</td>
 
     </tr>
 
     </tr>
 
     <tr>
 
     <tr>
 
       <td>Notes:</td>
 
       <td>Notes:</td>
−
       <td>NheI-HF;
+
       <td>Digest of gRNA-EndoA gBlock with NheI-HF and consequent heat inactivation 80°C. Samples were gibson ligated into p2ct_his_MBP_LBU_c2c2 and E.Coli NEB Turbo was transformed. Sample DNA was preped and digested to verify the correctness.
−
Heat inactivation 80°C
+
 
</td>
 
</td>
 
     </tr>
 
     </tr>
 
  <tr>
 
  <tr>
 
       <td>Results:</td>
 
       <td>Results:</td>
−
       <td></td>
+
       <td>resctriction digest showed false results; Thomas (supervisor) and Katja decided to redo the experiment</td>
 
     </tr>
 
     </tr>
 
</table>
 
</table>
  
−
<h3>4/24</h3>
+
<h4>Assembling p2ct_his_MBP_LBU_c2c2_gRNA-EndoA</h4>
−
<h4>Gibson ligation of Gblock EndoA in </h4>
+
<em>2018/05/16</em>
 
<table class="table table-borderless">
 
<table class="table table-borderless">
 
     <tr>
 
     <tr>
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     <tr>
 
     <tr>
 
       <td>Protocol:</td>
 
       <td>Protocol:</td>
−
       <td>restriction digest</td>
+
       <td>Gibson ligation, trafo, mini.prep, restriction digest</td>
 
     </tr>
 
     </tr>
 
     <tr>
 
     <tr>
 
       <td>Notes:</td>
 
       <td>Notes:</td>
−
       <td>
+
       <td>2x chemical transformation (turbo, dh5a) 1x electroporation (dh5a)
 
</td>
 
</td>
 
     </tr>
 
     </tr>
 
  <tr>
 
  <tr>
 
       <td>Results:</td>
 
       <td>Results:</td>
−
       <td></td>
+
       <td>some colonies. Sample DNA was prepped and digested to verify the plasmid. Restriction digest gave no correct bands for the plasmid. We suspected that the Cas13a template is not good anymore.</td>
 
     </tr>
 
     </tr>
 
</table>
 
</table>
  
−
<h3>4/24</h3>
+
<h4>Test digest of p2ct_his_MBP_LBU_c2c2_WT</h4>
−
<h4>Transformation of  p2ct_his_MBP_LBU_c2c2_WT_EndoA</h4>
+
<em>2018/05/28</em>
−
<table class="table table-borderless">
+
−
    <tr>
+
−
 
+
−
      <td>Participants:</td>
+
−
      <td>Katja</td>
+
−
    </tr>
+
−
    <tr>
+
−
      <td>Protocol:</td>
+
−
      <td>epo</td>
+
−
    </tr>
+
−
    <tr>
+
−
      <td>Notes:</td>
+
−
      <td>
+
−
</td>
+
−
    </tr>
+
−
<tr>
+
−
      <td>Results:</td>
+
−
      <td>colonies</td>
+
−
    </tr>
+
−
</table>
+
−
 
+
−
<h3>4/25</h3>
+
−
<h4>Mini-prep of  p2ct_his_MBP_LBU_c2c2_WT_EndoA</h4>
+
−
<table class="table table-borderless">
+
−
    <tr>
+
−
 
+
−
      <td>Participants:</td>
+
−
      <td>Katja</td>
+
−
    </tr>
+
−
    <tr>
+
−
      <td>Protocol:</td>
+
−
      <td>mini-prep</td>
+
−
    </tr>
+
−
    <tr>
+
−
      <td>Notes:</td>
+
−
      <td>
+
−
</td>
+
−
    </tr>
+
−
<tr>
+
−
      <td>Results:</td>
+
−
      <td></td>
+
−
    </tr>
+
−
</table>
+
−
 
+
−
<h3>4/25</h3>
+
−
<h4>Test digest of  p2ct_his_MBP_LBU_c2c2_WT_EndoA</h4>
+
−
<table class="table table-borderless">
+
−
    <tr>
+
−
 
+
−
      <td>Participants:</td>
+
−
      <td>Katja</td>
+
−
    </tr>
+
−
    <tr>
+
−
      <td>Protocol:</td>
+
−
      <td>restrictiondigest</td>
+
−
    </tr>
+
−
    <tr>
+
−
      <td>Notes:</td>
+
−
      <td>
+
−
</td>
+
−
    </tr>
+
−
<tr>
+
−
      <td>Results:</td>
+
−
      <td>did not work (PIC); we suspected that the cas13a plasmid is not good anymore</td>
+
−
    </tr>
+
−
</table>
+
−
 
+
−
<h3>4/26</h3>
+
−
<h4>Mini-prep of p2ct_his_MBP_LBU_c2c2_WT</h4>
+
−
<table class="table table-borderless">
+
−
    <tr>
+
−
 
+
−
      <td>Participants:</td>
+
−
      <td>Katja</td>
+
−
    </tr>
+
−
    <tr>
+
−
      <td>Protocol:</td>
+
−
      <td>mini-prep</td>
+
−
    </tr>
+
−
    <tr>
+
−
      <td>Notes:</td>
+
−
      <td>
+
−
</td>
+
−
    </tr>
+
−
<tr>
+
−
      <td>Results:</td>
+
−
      <td>92 ng/ul</td>
+
−
    </tr>
+
−
</table>
+
−
 
+
−
<h3>5/16</h3>
+
−
<h4>Restriction digest of p2ct_his_MBP_LBU_c2c2_WT</h4>
+
−
<table class="table table-borderless">
+
−
    <tr>
+
−
 
+
−
      <td>Participants:</td>
+
−
      <td>Katja</td>
+
−
    </tr>
+
−
    <tr>
+
−
      <td>Protocol:</td>
+
−
      <td>restriction digest</td>
+
−
    </tr>
+
−
    <tr>
+
−
      <td>Notes:</td>
+
−
      <td>NheI
+
−
</td>
+
−
    </tr>
+
−
<tr>
+
−
      <td>Results:</td>
+
−
      <td>92 ng/ul</td>
+
−
    </tr>
+
−
</table>
+
−
 
+
−
<h3>5/16</h3>
+
−
<h4>Gibson ligation of p2ct_his_MBP_LBU_c2c2_WT</h4>
+
−
<table class="table table-borderless">
+
−
    <tr>
+
−
 
+
−
      <td>Participants:</td>
+
−
      <td>Katja</td>
+
−
    </tr>
+
−
    <tr>
+
−
      <td>Protocol:</td>
+
−
      <td>Gibson ligation</td>
+
−
    </tr>
+
−
    <tr>
+
−
      <td>Notes:</td>
+
−
      <td>NheI
+
−
</td>
+
−
    </tr>
+
−
<tr>
+
−
      <td>Results:</td>
+
−
      <td>92 ng/ul</td>
+
−
    </tr>
+
−
</table>
+
−
 
+
−
<h3>5/16</h3>
+
−
<h4>Transformation of  p2ct_his_MBP_LBU_c2c2_WT_EndoA</h4>
+
−
<table class="table table-borderless">
+
−
    <tr>
+
−
 
+
−
      <td>Participants:</td>
+
−
      <td>Katja</td>
+
−
    </tr>
+
−
    <tr>
+
−
      <td>Protocol:</td>
+
−
      <td>chemisch, epo</td>
+
−
    </tr>
+
−
    <tr>
+
−
      <td>Notes:</td>
+
−
      <td>2x chemisch (turbo, dh5a) 1x epo (dh5a)
+
−
</td>
+
−
    </tr>
+
−
<tr>
+
−
      <td>Results:</td>
+
−
      <td>colonies</td>
+
−
    </tr>
+
−
</table>
+
−
 
+
−
<h3>5/16</h3>
+
−
<h4>Test digest of  p2ct_his_MBP_LBU_c2c2_WT_EndoA</h4>
+
−
<table class="table table-borderless">
+
−
    <tr>
+
−
 
+
−
      <td>Participants:</td>
+
−
      <td>Katja</td>
+
−
    </tr>
+
−
    <tr>
+
−
      <td>Protocol:</td>
+
−
      <td>restrictiondigest</td>
+
−
    </tr>
+
−
    <tr>
+
−
      <td>Notes:</td>
+
−
      <td>nheI
+
−
</td>
+
−
    </tr>
+
−
<tr>
+
−
      <td>Results:</td>
+
−
      <td>did not work; we suspected that the restrcition enzymes might not work or template is broken</td>
+
−
    </tr>
+
−
</table>
+
−
 
+
−
<h3>5/28</h3>
+
−
<h4>Test digest of  p2ct_his_MBP_LBU_c2c2_WT</h4>
+
 
<table class="table table-borderless">
 
<table class="table table-borderless">
 
     <tr>
 
     <tr>
Line 248: Line 63:
 
       <td>Notes:</td>
 
       <td>Notes:</td>
 
       <td>SalI NheI;
 
       <td>SalI NheI;
−
SalI;
 
−
NheI
 
−
 
 
</td>
 
</td>
 
     </tr>
 
     </tr>
Line 260: Line 72:
  
 
</div>
 
</div>
 +
</html>

Revision as of 15:57, 29 September 2018

Assembling p2ct_his_MBP_LBU_c2c2_gRNA-EndoA

2018/04/24 - 2018/04/26
Participants: Katja
Protocol: restriction digest, gibson ligation, chem trafo
Notes: Digest of gRNA-EndoA gBlock with NheI-HF and consequent heat inactivation 80°C. Samples were gibson ligated into p2ct_his_MBP_LBU_c2c2 and E.Coli NEB Turbo was transformed. Sample DNA was preped and digested to verify the correctness.
Results: resctriction digest showed false results; Thomas (supervisor) and Katja decided to redo the experiment

Assembling p2ct_his_MBP_LBU_c2c2_gRNA-EndoA

2018/05/16
Participants: Katja
Protocol: Gibson ligation, trafo, mini.prep, restriction digest
Notes: 2x chemical transformation (turbo, dh5a) 1x electroporation (dh5a)
Results: some colonies. Sample DNA was prepped and digested to verify the plasmid. Restriction digest gave no correct bands for the plasmid. We suspected that the Cas13a template is not good anymore.

Test digest of p2ct_his_MBP_LBU_c2c2_WT

2018/05/28
Participants: Katja
Protocol: restrictiondigest
Notes: SalI NheI;
Results: restriction enzymes and template are OK (PIC)