Team:ZJU-China/Processing

From 2014.igem.org

(Difference between revisions)
Line 10: Line 10:
<div class="zju_sec">
<div class="zju_sec">
-
<table class="img" style="float:none;width:100%">
+
<table class="img" style="float:none;width:100%;font-size:14pt;">
     <tr><th width="50%">Details in Socket E.coli</th><th width="50%">Tasks for you</th></tr>
     <tr><th width="50%">Details in Socket E.coli</th><th width="50%">Tasks for you</th></tr>
     <tr><td></td><td><p>Design circuit and find your parts DNA</p></td></tr>
     <tr><td></td><td><p>Design circuit and find your parts DNA</p></td></tr>

Revision as of 17:28, 16 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 rep-Int-exp.gif

Cell recovery after electrotranformation, liquid cultivation.

Int express after recombination, flip over attB/attP site

Stage switched, reporter 2 expressed 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 int+xis-flip lr.gif 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.