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Gel electrophoresis after colony PCR showed that most colonies merely contained the original dimer (around 1700), but two colonies did give good results (around 4200). The sequencing services of StarSEQ confirmed that the formation of a tetrameric 14-3-3 with GFP was finally successful.</h6><br/><br/> | Gel electrophoresis after colony PCR showed that most colonies merely contained the original dimer (around 1700), but two colonies did give good results (around 4200). The sequencing services of StarSEQ confirmed that the formation of a tetrameric 14-3-3 with GFP was finally successful.</h6><br/><br/> | ||
− | <h6><div class=" | + | <h6><div class="Figure_1"><img src="https://static.igem.org/mediawiki/2017/4/4c/T--TU-Eindhoven--14-3-3_tetramer_gel.png" width="500" height="500" /> |
<figcaption>Figure 1: 14-3-3 tetramer with GFP</figcaption></div></h6> | <figcaption>Figure 1: 14-3-3 tetramer with GFP</figcaption></div></h6> | ||
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+ | <h2>CT33 with Strep-tag</h2> | ||
+ | <h6><i>First approach</i><br> | ||
+ | We designed a gBlock encoding for an N-terminal Strep-tag II, followed by the mCherry fluorophore and the C-terminal CT33 sequence, in such a way that it can be placed into pBAD-DEST49 vector. This was performed using NEBuilder HiFi DNA Assembly Cloning Kit. With an expected length of approximately 1100 bp this proved successful in multiple colonies after colony PCR. This was also confirmed by sequencing.<br><br> | ||
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+ | <h6><div class="Figure_2"><img src="https://static.igem.org/mediawiki/2017/8/81/T--TU-Eindhoven--CT33_in_pBAD.png"> | ||
+ | <figcaption>Figure 1: 14-3-3 tetramer with GFP</figcaption></div></h6> | ||
+ | <br> | ||
Revision as of 13:37, 1 November 2017