Difference between revisions of "Template:BOKU-Vienna/GoldenGate iGEM Parts"
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<p>Because we worked during our work only with Golden Gate Systems, we had the problem how to send these parts to the iGEM - Registry. | <p>Because we worked during our work only with Golden Gate Systems, we had the problem how to send these parts to the iGEM - Registry. | ||
But we had an idea. What is if we combine Golden Gate Standard with the FC10 iGEM Standard?</p> | But we had an idea. What is if we combine Golden Gate Standard with the FC10 iGEM Standard?</p> | ||
| − | <p><br | + | <p><br></p> |
<h2 id="the-pcrs">The PCRs</h2> | <h2 id="the-pcrs">The PCRs</h2> | ||
<p>First of all, each Part and also the Vector pSB1C3 has to tranform into GoldenGate Standards with BbsI cut sites.</p> | <p>First of all, each Part and also the Vector pSB1C3 has to tranform into GoldenGate Standards with BbsI cut sites.</p> | ||
| − | <p><br | + | <p><br></p> |
<div class="row d-flex align-items-center"> | <div class="row d-flex align-items-center"> | ||
<div class="col-md-6"> | <div class="col-md-6"> | ||
<p>As you can see in the picture 1 we designed the primer, for each parts in this way that these have the cutsites for the igemregistry and also for BbsI.</p> | <p>As you can see in the picture 1 we designed the primer, for each parts in this way that these have the cutsites for the igemregistry and also for BbsI.</p> | ||
<p>The Vector pSB1C3 got also the Cutsite for BbsI in this way, that the BbsI recognition site was removed at the assembly. | <p>The Vector pSB1C3 got also the Cutsite for BbsI in this way, that the BbsI recognition site was removed at the assembly. | ||
| − | </div> | + | </p></div> |
<div class="col-md-6 col-image"> | <div class="col-md-6 col-image"> | ||
| − | <a href="/ | + | <a href="https://2018.igem.org/Team:BOKU-Vienna/pSB1C3iGEM1.jpg" data-title="Primer Design for GoldenGate Igem Registry" data-toggle="lightbox"> |
| − | <img src="/ | + | <img src="https://static.igem.org/mediawiki/2018/c/ce/T--BOKU-Vienna--2018_pSB1C3iGEM1.jpg" alt="Primer Design for GoldenGate Igem Registry"> |
</a> | </a> | ||
</div> | </div> | ||
<div class="col-md-4 col-image"> | <div class="col-md-4 col-image"> | ||
| − | <a href="/ | + | <a href="https://2018.igem.org/Team:BOKU-Vienna/gToffGfpSequenceiGEM1.jpg" data-title="Primer Design for GoldenGate Igem Registry" data-toggle="lightbox"> |
| − | <img src="/ | + | <img src="https://static.igem.org/mediawiki/2018/0/03/T--BOKU-Vienna--2018_gToffGfpSequenceiGEM1.jpg" alt="Primer Design for GoldenGate Igem Registry"> |
</a> | </a> | ||
</div> | </div> | ||
<div class="col-md-8"> | <div class="col-md-8"> | ||
<p>And also for each part the primer was designed in this way, that they contain the cutsites for BbsI, EcoRI, NotI, XbaI, SpeI and PstI. | <p>And also for each part the primer was designed in this way, that they contain the cutsites for BbsI, EcoRI, NotI, XbaI, SpeI and PstI. | ||
| − | </div> | + | </p></div> |
</div> | </div> | ||
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<p>The Method for each PCR was a standard method for Q5 (from NEB)</p> | <p>The Method for each PCR was a standard method for Q5 (from NEB)</p> | ||
<table class="table table-dark"> | <table class="table table-dark"> | ||
| − | <thead > | + | <thead> |
<tr><th> Name </th><th> Volume </th></tr> | <tr><th> Name </th><th> Volume </th></tr> | ||
</thead> | </thead> | ||
Revision as of 09:08, 4 October 2018
Contents
Golden Gate iGEM Parts
Send iGEM-Parts easyer and faster to the iGEM-Registry.
Because we worked during our work only with Golden Gate Systems, we had the problem how to send these parts to the iGEM - Registry. But we had an idea. What is if we combine Golden Gate Standard with the FC10 iGEM Standard?
The PCRs
First of all, each Part and also the Vector pSB1C3 has to tranform into GoldenGate Standards with BbsI cut sites.
As you can see in the picture 1 we designed the primer, for each parts in this way that these have the cutsites for the igemregistry and also for BbsI.
The Vector pSB1C3 got also the Cutsite for BbsI in this way, that the BbsI recognition site was removed at the assembly.
<a href="" data-title="Primer Design for GoldenGate Igem Registry" data-toggle="lightbox"> <img src="
" alt="Primer Design for GoldenGate Igem Registry"> </a>
<a href="" data-title="Primer Design for GoldenGate Igem Registry" data-toggle="lightbox"> <img src="
" alt="Primer Design for GoldenGate Igem Registry"> </a>
And also for each part the primer was designed in this way, that they contain the cutsites for BbsI, EcoRI, NotI, XbaI, SpeI and PstI.
PCR Method
The Method for each PCR was a standard method for Q5 (from NEB)
| Name | Volume |
|---|---|
| Q5 High-Fidelity 2X Master Mix | 25 µL |
| 10 µM Forward Primer | 2.5 µL |
| 10 µM Reverse Primer | 2.5 µL |
| Template DNA | 2 µL |
| Nuclease free water | 18 µL |