Difference between revisions of "Template:Team:TU Eindhoven/Timeline HTML"

Line 45: Line 45:
 
<h4>Week 28</h4>
 
<h4>Week 28</h4>
 
<h3> - Et tu, pET-Duet? </h3>
 
<h3> - Et tu, pET-Duet? </h3>
 +
<br \>
 
<span class="activiteit"> Gibson Assembly:</span>
 
<span class="activiteit"> Gibson Assembly:</span>
 
<ul class="activiteitlijst">
 
<ul class="activiteitlijst">
 
<li><span class="activiteit">Linearizing pET-Duet-1 for Gibson Assembly and debugging </span></li>
 
<li><span class="activiteit">Linearizing pET-Duet-1 for Gibson Assembly and debugging </span></li>
−
<li><span class="activiteit">Amplification of the inserts for traditional cloning</span></li>
 
−
<li><span class="activiteit">Xbal &amp; Pstl digestion of the pET-Duet-1 vector (MCS-1) for traditional cloning</span></li>
 
 
</ul>
 
</ul>
 
<br \>
 
<br \>
 
<span class="activiteit"> Traditional cloning:</span>
 
<span class="activiteit"> Traditional cloning:</span>
 
<ul class="activiteitlijst">
 
<ul class="activiteitlijst">
−
<li><span class="activiteit">Xba1 &amp; Pst1 digestion of the inserts (MCS-1) for traditional cloning</span></li>
+
<li><span class="activiteit">Amplification of the inserts</span></li>
−
<li><span class="activiteit">Nde1 &amp; Kpn1 digestion of the inserts (MCS-2) for traditional cloning</span></li>
+
<li><span class="activiteit">Xbal &amp; Pstl digestion of the pET-Duet-1 vector (MCS-1)</span>
 +
<li><span class="activiteit">Xba1 &amp; Pst1 digestion of the inserts (MCS-1)</span></li>
 +
<li><span class="activiteit">Nde1 &amp; Kpn1 digestion of the inserts (MCS-2)</span></li>
 
<li><span class="activiteit">Ligation</span></li>
 
<li><span class="activiteit">Ligation</span></li>
 +
<li><span class="activiteit">Transformation in NB</span></li>
 +
<li><span class="activiteit">Colony PCR &amp; gel electrophorese: The inserts are succesfully ligated</span></li>
 +
<li><span class="activiteit">Culturing of the colonies with the correct plasmid</span></li>
 +
<li><span class="activiteit">Making a glycerol stock & sending the DNA for sequencing</span></li>
 
</ul>
 
</ul>
  

Revision as of 07:32, 10 July 2015





Timeline



Week 26

- Preparing for take-off


  • Inventory of the lab supplies
  • Pouring LB Agar plates
  • Amplification of the pET-Duet-1 vector
  • Assessment of the safety requirements
  • Preparing stocks for antibiotics, glycerol, LB & MilliQ


Week 27

- The Clone Wars


Gibson Assembly:
  • Linearizing pET-Duet-1 for Gibson Assembly
  • Our first Gibson Assembly!

Traditional cloning:
  • Amplification of the pET-Duet-1 vector
  • Nde1 & Kpn1 digestion of the pET-Duet-1 vector (MCS-2) for traditional cloning


Week 28

- Et tu, pET-Duet?


Gibson Assembly:
  • Linearizing pET-Duet-1 for Gibson Assembly and debugging

Traditional cloning:
  • Amplification of the inserts
  • Xbal & Pstl digestion of the pET-Duet-1 vector (MCS-1)
  • Xba1 & Pst1 digestion of the inserts (MCS-1)
  • Nde1 & Kpn1 digestion of the inserts (MCS-2)
  • Ligation
  • Transformation in NB
  • Colony PCR & gel electrophorese: The inserts are succesfully ligated
  • Culturing of the colonies with the correct plasmid
  • Making a glycerol stock & sending the DNA for sequencing