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Team:ZJU-China/Processing
From 2014.igem.org
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| + | <b><big><big><big><big><center>Working process of GeneSocket</center></big></big></big></big></b><br /> | ||
| + | Look through this page, you will understand the working process of GeneSocket inside the cell and learn corresponding experiment step. | ||
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Revision as of 00:40, 18 October 2014
| Team Sponsor | Contact us | ||
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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 |
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| Powered by Media Wiki. Zhejiang University. | Email Address: | zjuchina2014 @ 163.com | |
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. |
