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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<table border=1px class="img" cellspacing="0" bordercolorlight="#333333" bordercolordark="#efefef" style="float:none;width:100%;font-size:14pt;" >
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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 bgcolor=#eafeff><td></td><td><p>Design circuit and find your parts DNA</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 bgcolor=#eafeff><td></td><td><p>PCR to add Homeoregion, Ligase standard parts.</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 bgcolor=#eafeff><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>Inserted circuit recombine biterminator unit<br>
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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 bgcolor=#eafeff><td><p>Int express after recombination, flip over attB/attP site</p></td></tr>
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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"/>
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</p></td><td><p>plate cultivation, Use reporter 2 to select candidate cells</p></td></tr>
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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/2014/6/6b/Zju-int-xis-flip-lr.gif " width="270" height="200" align ="center" style="float:none"/>
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<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" style="float:none"/>
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</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 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 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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<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
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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

Team SponsorContact us

Zhejiang University ZJU International Education Fundation College of Life Sciences ZJU School of Medicine Zhejiang University

College of Agriculture & Biotechnology ZJU Faculty of Science Zhejiang University Undergraduate School of ZJU Department of Polymer Sciencce and Engneering Zhejiang University

International Relations Zhejiang University Vazyme Nanking ™ Co.

 Team Forum(BBS): ZJU-China 2014 Team Forum on www.zjubiolab.zju.edu.cn
Post Address: Biolab Center Room 413, ZJU Zijin'gang Campus,
Yuhang Tang Road No.866, Hangzhou, Zhejiang
Powered by . Zhejiang University. Email Address: zjuchina2014 @ 163.com

 

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

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