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 | ||
- | [[File: | + | [[File:zju_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 | ||
- | [[File: | + | [[File:zju_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 | ||
- | [[File: | + | [[File:zju_int_xis_flip_lr.gif]] |
- | [[File: | + | [[File:zju_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> |
Revision as of 09:42, 17 October 2014
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 [[File:zju_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:zju_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:zju_int_xis_flip_lr.gif]] [[File:zju_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. |