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</figure> | </figure> | ||
<figure> | <figure> | ||
− | <figure class="makeresponsive" style="width: | + | <figure class="makeresponsive" style="width: 70%; float: left;"> |
<img src="https://static.igem.org/mediawiki/2017/4/4d/EvaluationVectorCloning.png" alt="A scheme explaining the cloning of the Evaluation Vector."> | <img src="https://static.igem.org/mediawiki/2017/4/4d/EvaluationVectorCloning.png" alt="A scheme explaining the cloning of the Evaluation Vector."> | ||
− | <figcaption><b>Figure 2: Cloning scheme of the Evaluation Vector.</b> The Detailed cloning workflow which led to the finished Evaluation Vector construct with the pBS1C backbone.</figcaption></figure>Finally, we combined our new MCS, by ligating the digested RFPsyn2 (cut with XbaI and AgeI) with the <i>lacZα</i> fragment (cut with AgeI and PstI). This MCS was inserted into the pBS1C-P<sub>xylA</sub> backbone, which was prior opened using BsaI (resulting in an XbaI overhang) and PstI (Figure 2). The final construct of our EV was verified by sequencing.<br><br>We decided to additionally also provide this MCS as a BioBrick. Therefore we cloned it into the pSB1C3 backbone (via EcoRI and PstI digest) and verified the construct by sequencing. It has been submitted to the parts registry under <a target="_blank" href ="http://parts.igem.org/Part:BBa_K2273107">BBa_K2273107</a>.<br><br>Detailed methods can be found in our <a target="_blank" href ="https://2017.igem.org/Team:TU_Dresden/Experiments">protocol collection section</a>. All primers used can be found in our primer collection table. | + | <figcaption><b>Figure 2: Cloning scheme of the Evaluation Vector.</b> The Detailed cloning workflow which led to the finished Evaluation Vector construct with the pBS1C backbone.</figcaption></figure>Finally, we combined our new MCS, by ligating the digested RFPsyn2 (cut with XbaI and AgeI) with the <i>lacZα</i> fragment (cut with AgeI and PstI). This MCS was inserted into the pBS1C-P<sub>xylA</sub> backbone, which was prior opened using BsaI (resulting in an XbaI overhang) and PstI (Figure 2). The final construct of our EV was verified by sequencing.<br><br>We decided to additionally also provide this MCS as a BioBrick. Therefore we cloned it into the pSB1C3 backbone (via EcoRI and PstI digest) and verified the construct by sequencing. It has been submitted to the parts registry under <a target="_blank" href ="http://parts.igem.org/Part:BBa_K2273107">BBa_K2273107</a>.<br><br>Detailed methods can be found in our <a target="_blank" href ="https://2017.igem.org/Team:TU_Dresden/Experiments">protocol collection section</a>. All primers used can be found in our primer collection table. |
</figure> | </figure> | ||
+ | <a class="pdf-resources" href="https://2017.igem.org/File:T--TU_Dresden--M_Primerlist.pdf">Primerlist</a> | ||
</div> | </div> | ||
Revision as of 14:17, 24 October 2017