Team:WHU-China/templates/standardpage results

From 2013.igem.org

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<div class="mac" style="height:autopx;width:67%;float:right;margin-right:7%;*width:98%;*border: 3px solid green;">
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<h1 style="width:100%;font-size:20px;">title</h1></br>
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<h1 style="width:100%;font-size:20px;">Current Result</h1></br>
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<b>Double Promoter</b></br>
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<img src="https://static.igem.org/mediawiki/2013/9/92/WHUlyDP.png" /></br>
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We have successfully ligate tandem double promoters with J23102 J23106 and J23116 in different orders: J23102-J23102, J23102-J23106, J23106-J23102, J23116-J23102, J23106-J23106, J23106-J23116, J23116-J23106. And we have submitted these new biobrick. Unfortunately, other combination failed because of mutation showed by sequence result.</br>
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And we have detected the transcription strength of these tandem double promoters.</br>
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</br></br>
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<b>Guide RNA</b></br>
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<img src="https://static.igem.org/mediawiki/2013/1/14/WHUlyTandem_Promoters.JPG" /></br>
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We chose tandem double promoters J23106-J23116 as our target. Based on this, we designed serval targeting sites, including both activation(A1,A2,A3,A4,A5) and repression(R1, R2) sites after searching the PAM region. And through a three-cycle overlap PCR, we construct these functinal guide RNA.</br>
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<div style="width:100%;text-align:center">
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<img src="https://static.igem.org/mediawiki/2013/5/5f/WHUlyRNA1.png" /></br>
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Here shows the agarose electrophoresis:</br>
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<img src="https://static.igem.org/mediawiki/2013/3/32/WHUlyGRNA%E6%89%A9%E5%A2%9E%E7%BB%93%E6%9E%9C.jpg" />
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<img src="https://static.igem.org/mediawiki/2013/d/d7/WHUlyN20.JPG" /></br>
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</div>
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</br></br>
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<b>dCas9 Devices</b></br>
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First, we add dCas9 gene( amplify through PCR) to express vector with promoter to construct a new biobrick. We name this new biobrick BBa_K1081000.</br>
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<div style="width:100%;text-align:center">
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<img src="https://static.igem.org/mediawiki/2013/1/14/WHUlyTandem_Promoters.JPG" /></br>
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</div>
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Revision as of 19:42, 27 September 2013

Current Result


Double Promoter

We have successfully ligate tandem double promoters with J23102 J23106 and J23116 in different orders: J23102-J23102, J23102-J23106, J23106-J23102, J23116-J23102, J23106-J23106, J23106-J23116, J23116-J23106. And we have submitted these new biobrick. Unfortunately, other combination failed because of mutation showed by sequence result.
And we have detected the transcription strength of these tandem double promoters.


Guide RNA

We chose tandem double promoters J23106-J23116 as our target. Based on this, we designed serval targeting sites, including both activation(A1,A2,A3,A4,A5) and repression(R1, R2) sites after searching the PAM region. And through a three-cycle overlap PCR, we construct these functinal guide RNA.

Here shows the agarose electrophoresis:



dCas9 Devices
First, we add dCas9 gene( amplify through PCR) to express vector with promoter to construct a new biobrick. We name this new biobrick BBa_K1081000.


subtitle

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