Team:Peking/Notebook
Notebook
Week 1
26/06/2017-02/07/2017Design primers.
Construct standard structure of integrase int5 and int8.
Week 2
03/07/2017-09/07/2017Mutate integrase int8, int10, p22 for Golden Gate Assembly.
Construct the expression system of integrase TP901, int2, int3, int4, int5, int7, int8, p21, FimB, FimE, HbiF, Vcre, Scre, Cre, Dre, Vika, Lambda-int.
Construct the reporter system utilized the structure of attB-Terminator-attP.
Week 3
10/07/2017-16/07/2017Mutate Bxb1-gp35, phiC31 for Golden Gate Assembly.
Ask for some plasmids of repressilator.
Week 4
17/07/2017-23/07/2017Construct the reporter system of Bxb1, TP901, phiC31.
Prepare for repressilator test.
Week 5
24/07/2017-30/07/2017Construct the expression system of integrase TP901, Bxb1-gp35, phiC31 without RBS.
Design Microfluidic device for the oscillation test of repressilator.
Week 6
31/07/2017-06/08/2017Construct the expression system of integrase TP901, Bxb1-gp35 with a series RBS of different transcription initiate rate.
Manufacture seven-segment display microfluidic device.
Week 7
07/08/2017-13/08/2017Construct the expression system of integrase TP901, Bxb1-gp35 with another series RBS of different transcription initiate rate.
Image period of LP117 oscillation around 15 hours by microfluidic device.
Week 8
14/08/2017-20/08/2017Construct the expression system of integrase phiC31 with a series RBS of different transcription initiate rate.
Manufacture Sand Clock microfluidic device.
Prepare for parts construction.
Week 9
21/08/2017-27/08/2017Construct the expression system and reporter system of integrase-RDF (recombination directionality factor) fusion protein.
Test integrase recombination efficiency of TP901, Bxb1-gp35, phiC31 by Flow Cytometer.
Image period of plpt20 oscillation around 7 hours.
Week 10
28/08/2017-03/09/2017Make adjustment for the expression system of integrase TP901, Bxb1-gp35, phiC31 to improve the recombination efficiency, and rebuild the expression system.
Test the adjusted integrase recombination efficiency of TP901, Bxb1-gp35, phiC31 by Flow Cytometer.
Parts construction of integrase TP901.
Site mutation of plpt20 original plasmid.
Week 11
04/09/2017-10/09/2017Construct the front end of Flip-Flop.
Construct the expression system of integrase-RDF (recombination directionality factor) fusion protein.
Site mutation of plpt20 original plasmid.
Week 12
11/09/2017-17/09/2017Make adjustment again for the expression system of integrase TP901, Bxb1-gp35, phiC31 to improve the recombination efficiency, and rebuild the expression system.
Test the adjusted integrase recombination efficiency of TP901, Bxb1-gp35, phiC31 by Flow Cytometer.
Construct the reporter system of integrase-RDF (recombination directionality factor) fusion protein.
Parts construction of integrase int2, int5.
Construct the expression system for oscillator plpt20 with integrase Bxb1-gp35.
Week 13
18/09/2017-24/09/2017Construct TP901, Bxb1, phiC31 expression system of integrase-RDF.
Parts construction of integrase expression system.
Construct the expression system for oscillator plpt20 with integrase T7 promoter.
Week 14
25/09/2017-01/10/2017Test the recombination efficiency of TP901, Bxb1 integrase-RDF by Flow Cytometer.
Parts construction of integrase reporter system.
Week 15
02/10/2017-08/10/2017Make adjustment for integrase-RDF expression system to improve recombination efficiency.
Test the recombination efficiency of adjusted integrase-RDF.
Parts construction of integrase expression system and reporter system.
Construct reporter system for oscillator plpt41 with T7 promoter, Bxb1-gp35.
Image repressilator-drived DNA recombination.
Week 16
09/10/2017-15/10/2017Measure the state transition efficiency of the Forward latch.
Measure the recombination efficiency of the excisionases.
Parts construction of the integrases and excisionases reporter systems.
Measure the flipping efficiency and filter efficiency of the terminators flanked by a pair of attB and attP sites.
Week 17
16/10/2017-22/10/2017Measure the recombination efficiency of the excisionases.
Parts construction of the integrases and excisionases reporter systems.
Measure the flipping efficiency and filter efficiency of the terminators flanked by two pair of attB and attP sites.
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