Team:ZJU-China/Processing

From 2014.igem.org

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     <tr><td><p>Inserted circuit(dsDNA) are introduced into Socket.coli.</p></td><td><p>electrotranformation</p></td></tr>
     <tr><td><p>Inserted circuit(dsDNA) are introduced into Socket.coli.</p></td><td><p>electrotranformation</p></td></tr>
     <tr><td><p>Inserted circuit recombine biterminator unit
     <tr><td><p>Inserted circuit recombine biterminator unit
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[[file:rep-Int-exp.gif]]</p></td><td rowspan="2"><p>Cell recovery after electrotranformation, liquid cultivation.</p></td></tr>
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[[File:rep-Int-exp.gif]]</p></td><td rowspan="2"><p>Cell recovery after electrotranformation, liquid cultivation.</p></td></tr>
     <tr><td><p>Int express after recombination, flip over attB/attP site</p></td></tr>
     <tr><td><p>Int express after recombination, flip over attB/attP site</p></td></tr>
     <tr><td><p>Stage switched, reporter 2 expressed
     <tr><td><p>Stage switched, reporter 2 expressed
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[[file:int-flip-bp.gif]]
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[[File:int-flip-bp.gif]]
</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>
     <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
     <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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[[file:int+xis-flip lr.gif]]
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[[File:int+xis-flip lr.gif]]
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[[file:int+xis-flip T.gif]]
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[[File:int+xis-flip T.gif]]
</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>
     <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>
     <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>
     <tr><td></td><td><p>If circuit construction is over, cultivate cell in 42℃ to discard Support device.</p></td></tr>
     <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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[[File:full cartoon.mov]]
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Revision as of 09:04, 17 October 2014

 

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 [[File:rep-Int-exp.gif]]

Cell recovery after electrotranformation, liquid cultivation.

Int express after recombination, flip over attB/attP site

Stage switched, reporter 2 expressed [[File:int-flip-bp.gif]]

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 [[File:int+xis-flip lr.gif]] [[File:int+xis-flip T.gif]]

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.

[[File:full cartoon.mov]]