Team:ZJU-China/Processing

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Revision as of 23:56, 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

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.