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<br> | <tr><td><p>Inserted circuit recombine biterminator unit<br> | ||
- | <img src="https://static.igem.org/mediawiki/2014/ | + | <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> |
<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 | ||
- | <img src="https://static.igem.org/mediawiki/2014/ | + | <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> | ||
<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 | ||
- | <img src="https://static.igem.org/mediawiki/2014/ | + | <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> </p><p> </p> | <p> </p><p> </p> | ||
<img src="https://static.igem.org/mediawiki/2014/2/2b/ZJU_Teminator_flipover.gif" width="270" height="200" align ="center" style="float:none"/> | <img src="https://static.igem.org/mediawiki/2014/2/2b/ZJU_Teminator_flipover.gif" width="270" height="200" align ="center" style="float:none"/> |
Revision as of 01:33, 18 October 2014
Look through this page, you will understand the working process of GeneSocket inside the cell and learn corresponding experiment step.
Details in Socket E.coli | Tasks 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. |