"
Team:ZJU-China/Processing
From 2014.igem.org
| Team Sponsor | Contact us | ||
|---|---|---|---|
|
|
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 Media Wiki. Zhejiang University. | Email Address: | zjuchina2014 @ 163.com | |
| 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: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. |
