Team:BostonU/Results

From 2012.igem.org

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<h9>MoClo level 0 Vectors and Parts</h9>
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<h9>Creating Destination Vectors</h9>
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<br><h7>Below are the step by step results we obtained  in making all of our destination vectors:<p>
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<br><img src="https://static.igem.org/mediawiki/2012/e/e3/Sum1.png" width="300px">lacz in DV0 <br>
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<li>We first transformed the destination vectors  into cells on to plates with IPTG/X-Gal for blue white screening. In this case,  we wanted to see blue cells, indicative of the presence of LacZ. Our  destination vectors also varied in whether the backbone is phosphorylated or  dephosphorylated. In this picture, the dephosphorylated showed higher  transformation efficiency by preventing recircularization of the backbone,  yielding more cells transformed with the LacZ as part of the destination  vector.</li>
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<img src="https://static.igem.org/mediawiki/2012/a/ac/Sum2.png" width="300px">dv1 w/lacz <br>
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<img src="https://static.igem.org/mediawiki/2012/a/ac/Sum2.png" width="300px">
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<li>We checked whether the destination vectors contained the LacZ fragment amplified with the moclo fusion sites by running digest of cut vs uncut. In the gel picture, each SM refers to a different destination vector and the C and UC means cut and uncut, respectively. <li>
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<br><img src="https://static.igem.org/mediawiki/2012/e/e3/Sum1.png" width="300px">
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<li>The final step is sequencing the destination vectors to confirm the correct orientation of LacZ. In the nucleotide sequence file and the complementary diagram, the LacZ sequence, 2 fusion sites and restriction sites are highlighted in different colors. The trace file shows confidence of the sequencing.<li>
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<img src="https://static.igem.org/mediawiki/2012/a/ac/Sum2.png" width="300px">
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<img src="https://static.igem.org/mediawiki/2012/thumb/d/da/Sum3.png/800px-Sum3.png" width="300px">seq veri dv1<br>
<img src="https://static.igem.org/mediawiki/2012/thumb/d/da/Sum3.png/800px-Sum3.png" width="300px">seq veri dv1<br>
<img src="https://static.igem.org/mediawiki/2012/0/04/Sum4.png" width="300px"> seq veri dv1 finch<br>
<img src="https://static.igem.org/mediawiki/2012/0/04/Sum4.png" width="300px"> seq veri dv1 finch<br>

Revision as of 16:51, 3 October 2012

BostonU iGEM Team: Welcome


Results Summary


Creating Destination Vectors
Below are the step by step results we obtained in making all of our destination vectors:

  • We first transformed the destination vectors into cells on to plates with IPTG/X-Gal for blue white screening. In this case, we wanted to see blue cells, indicative of the presence of LacZ. Our destination vectors also varied in whether the backbone is phosphorylated or dephosphorylated. In this picture, the dephosphorylated showed higher transformation efficiency by preventing recircularization of the backbone, yielding more cells transformed with the LacZ as part of the destination vector.
  • We checked whether the destination vectors contained the LacZ fragment amplified with the moclo fusion sites by running digest of cut vs uncut. In the gel picture, each SM refers to a different destination vector and the C and UC means cut and uncut, respectively.

  • The final step is sequencing the destination vectors to confirm the correct orientation of LacZ. In the nucleotide sequence file and the complementary diagram, the LacZ sequence, 2 fusion sites and restriction sites are highlighted in different colors. The trace file shows confidence of the sequencing.
  • seq veri dv1
    seq veri dv1 finch
    reg pcr
    moclo 0 comp
    ligationpcr
      MoClo level 0 Vectors and Parts

      Things to include: gels, sequence data, plate pictures level 1 moclo l0 moclo gene l0 moclo term l0 moclo rbs l0 moclo promotor


    Basic Genetic Circuits

      Things to include: SBOL figures of circuits, gels, sequence data, plate pictures, characterization data summary

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