Team:ZJU-China/Processing

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<b><big><big><big><big><center>Working process of GeneSocket</center></big></big></big></big></b><br />
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<p style="font-size:13pt">Look through this page, and you will understand the working process of GeneSocket inside the cell and learn corresponding experiment step.</p>
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     <tr><th width="50%">Details in Socket E.coli</th><th width="50%">Tasks for you</th></tr>
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     <tr><td></td><td><p>Design circuit and find your parts DNA</p></td></tr>
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     <tr bgcolor=#cccccc><th width="50%">Details in Socket E.coli</th><th width="50%">Tasks for you</th></tr>
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     <tr><td></td><td><p>Use GS-BOX to design the strategy of circuit construction</p></td></tr>
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     <tr bgcolor=#eafeff><td></td><td><p>Design circuit and find your parts DNA</p></td></tr>
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     <tr><td></td><td><p>PCR to add Homeoregion, Ligase standard parts.</p></td></tr>
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     <tr bgcolor=#eafeff><td></td><td><p>Use GS-BOX to design the strategy of circuit construction</p></td></tr>
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     <tr><td><p>Lambda red protein is expressed during cultivation</p></td><td><p>Making electrotranformation competent cells, use reporter 1 to select.</p></td></tr>
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     <tr bgcolor=#eafeff><td></td><td><p>PCR to add Homeoregion, Ligase standard parts.</p></td></tr>
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     <tr><td><p>Inserted circuit(dsDNA) are introduced into Socket.coli.</p></td><td><p>electrotranformation</p></td></tr>
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     <tr bgcolor=#eafeff><td><p>Lambda red protein is expressed during cultivation</p></td><td><p>Making electrotranformation competent cells, use reporter 1 to select.</p></td></tr>
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     <tr><td><p>Inserted circuit recombine biterminator unit<br>
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     <tr bgcolor=#eafeff><td><p>Inserted circuit(dsDNA) are introduced into Socket.coli.</p></td><td><p>electrotranformation</p></td></tr>
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<img src="https://static.igem.org/mediawiki/2014/3/3b/Zju_rep_Int_exp.gif" width="270" height="200" align ="center"/></p></td><td rowspan="2"><p>Cell recovery after electrotranformation, liquid cultivation.</p></td></tr>
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     <tr bgcolor=#eafeff><td><p>Inserted circuit recombine biterminator unit<br>
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     <tr><td><p>Int express after recombination, flip over attB/attP site</p></td></tr>
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<img src="https://static.igem.org/mediawiki/2014/4/4c/Zju-rep-Int-exp.gif" width="270" height="200" align ="center" style="float:none"/></p></td><td rowspan="2"><p>Cell recovery after electrotranformation, liquid cultivation.</p></td></tr>
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     <tr><td><p>Stage switched, reporter 2 expressed
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     <tr bgcolor=#eafeff><td><p>Int express after recombination, flip over attB/attP site</p></td></tr>
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<img src="https://static.igem.org/mediawiki/2014/7/74/ZJU_int_flip_bp.gif" width="270" height="200" align ="center"/>
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     <tr bgcolor=#eafeff><td><p>Stage switched, reporter 2 expressed
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<img src="https://static.igem.org/mediawiki/2014/b/bc/ZJU-int-flip-bp.gif" width="270" height="200" align ="center" style="float:none"/>
</p></td><td><p>plate cultivation, Use reporter 2 to select candidate cells</p></td></tr>
</p></td><td><p>plate cultivation, Use reporter 2 to select candidate cells</p></td></tr>
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     <tr><td><p>Xis express, Int+Xis will flip over attL/attR sites. Stage switched again and inverted biterminator unit flipped over to silence Int expression
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     <tr bgcolor=#eafeff><td><p>Xis express, Int+Xis will flip over attL/attR sites. Stage switched again and inverted biterminator unit flipped over to silence Int expression
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<img src="https://static.igem.org/mediawiki/2014/4/4c/Zju_int_xis_flip_lr.gif " width="270" height="200" align ="center"/>
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<img src="https://static.igem.org/mediawiki/2014/6/6b/Zju-int-xis-flip-lr.gif " width="270" height="200" align ="center" style="float:none"/>
<p>&nbsp;&nbsp;</p><p>&nbsp;&nbsp;</p>
<p>&nbsp;&nbsp;</p><p>&nbsp;&nbsp;</p>
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<img src="https://static.igem.org/mediawiki/2014/2/2b/ZJU_Teminator_flipover.gif" width="270" height="200" align ="center"/>
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<img src="https://static.igem.org/mediawiki/2014/2/2b/ZJU_Teminator_flipover.gif" width="270" height="200" align ="center" style="float:none"/>
</p></td><td><p><b>select positive colony to amplify, Arabinose induce at the main time. Use plasmid backbone resistance to select.</b></p></td></tr>
</p></td><td><p><b>select positive colony to amplify, Arabinose induce at the main time. Use plasmid backbone resistance to select.</b></p></td></tr>
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     <tr><td></td><td><p>plate cultivation, Use reporter 1 to select candidate cells. This survival cells are ready for the next recombination round.</p></td></tr>
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     <tr bgcolor=#eafeff><td></td><td><p>plate cultivation, Use reporter 1 to select candidate cells. This survival cells are ready for the next recombination round.</p></td></tr>
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     <tr><td></td><td><p>If circuit construction is over, cultivate cell in 42℃ to discard Support device.</p></td></tr>
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     <tr bgcolor=#eafeff><td></td><td><p>If circuit construction is over, cultivate cell in 42℃ to discard Support device.</p></td></tr>
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        <td><img  src="https://static.igem.org/mediawiki/2014/4/47/ZJU_left_arow.png"> </img></td><td> <a href="https://2014.igem.org/Team:ZJU-China/GeneSocket">Previous: GeneSocket</a></td>
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        <td><a href="https://2014.igem.org/Team:ZJU-China/Design">Next: Circuit Design</a> </td><td><img  src="https://static.igem.org/mediawiki/2014/1/19/ZJU_right_arow.png" > </img> </td>
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Latest revision as of 03:36, 18 October 2014

 

Working process of GeneSocket

Look through this page, and you will understand the working process of GeneSocket inside the cell and learn corresponding experiment step.

Details in Socket E.coliTasks for you

Design circuit and find your parts DNA

Use GS-BOX to design the strategy of circuit construction

PCR to add Homeoregion, Ligase standard parts.

Lambda red protein is expressed during cultivation

Making electrotranformation competent cells, use reporter 1 to select.

Inserted circuit(dsDNA) are introduced into Socket.coli.

electrotranformation

Inserted circuit recombine biterminator unit

Cell recovery after electrotranformation, liquid cultivation.

Int express after recombination, flip over attB/attP site

Stage switched, reporter 2 expressed

plate cultivation, Use reporter 2 to select candidate cells

Xis express, Int+Xis will flip over attL/attR sites. Stage switched again and inverted biterminator unit flipped over to silence Int expression

  

  

select positive colony to amplify, Arabinose induce at the main time. Use plasmid backbone resistance to select.

plate cultivation, Use reporter 1 to select candidate cells. This survival cells are ready for the next recombination round.

If circuit construction is over, cultivate cell in 42℃ to discard Support device.

                            

Previous: GeneSocket Next: Circuit Design