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Team:Peking S/lab/notebook/xjy
From 2011.igem.org
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| - | == ''' | + | == '''Summary''' == |
| - | Mainly in charge of microfluidic device, I | + | Mainly in charge of microfluidic device, I created a bilayer microfluidic system for our project, which provides a suitable space for different cells to be spatially separated apart and population controlled, as well as allowing chemical wires to transfer freely from each side to the other. What’s more, I also take part in molecular cloning of TPP regulator, characterization of chemical wire toolbox, and modeling work of XOR gate. |
=='''Contents'''== | =='''Contents'''== | ||
| Line 62: | Line 62: | ||
[<html><a href="#top">TOP</a></html>] | [<html><a href="#top">TOP</a></html>] | ||
===6.25=== | ===6.25=== | ||
| + | Design and draw blueprint of the first chip. | ||
===6.26=== | ===6.26=== | ||
| + | Do the photoetching of the first chip and mould the PDMS. | ||
===6.27=== | ===6.27=== | ||
| + | Make chips and test under microscope. Pick the relatively suitable size of chamber. | ||
===6.28=== | ===6.28=== | ||
| + | Design and draw blueprint of the second chip. | ||
===6.29=== | ===6.29=== | ||
| + | Do the photoetching of the second chip and mould the PDMS. | ||
===6.30=== | ===6.30=== | ||
| + | Make chips and test under microscope. | ||
==July== | ==July== | ||
| Line 134: | Line 140: | ||
[<html><a href="#top">TOP</a></html>] | [<html><a href="#top">TOP</a></html>] | ||
===7.1=== | ===7.1=== | ||
| + | Change the flow velocity and test the chip under microscope, take some photos of the cells in the chamber, mould new chips. | ||
===7.2=== | ===7.2=== | ||
| + | Make chips and test under microscope with slowed flow velocity, shoot a movie of the growing population in a chamber,and mould new chips. | ||
===7.3=== | ===7.3=== | ||
| + | Test under microscope with slowed flow velocity and add a heater (37℃) to the system, the chip works well. Mould new chips. | ||
===7.4=== | ===7.4=== | ||
| + | Make chips and discussion the new chip. | ||
===7.5=== | ===7.5=== | ||
| + | Design and draw blueprint of the third chip,which has wider channels and two kinds of chamber sizes. | ||
===7.6=== | ===7.6=== | ||
| + | Do the photoetching of the third chip and mould the PDMS. | ||
===7.7=== | ===7.7=== | ||
| + | Try the two-layered chips with a membrane beside each side of chips. | ||
===7.8=== | ===7.8=== | ||
| + | Try to use photoetching machine to align the markers on the chips. | ||
===7.9=== | ===7.9=== | ||
| + | Try to use common microscope to align the markers on the chips, by hands, successfully. Take photos of both sides (RFP and GFP) under fluorescence microscope. | ||
===7.10=== | ===7.10=== | ||
| + | Design and draw blueprint of the fourth chip,with longer channels. | ||
| + | |||
| + | ===7.11=== | ||
| + | Do the ligation of PBAD_AfsA, Pc_ArpA, Pqrr_gfp. | ||
===7.12=== | ===7.12=== | ||
| + | Do the ligation of PBAD_AfsA, Pc_ArpA, Pqrr_gfp. | ||
===7.13=== | ===7.13=== | ||
| + | Do the ligation of PBAD_AfsA, Pc_ArpA, Pqrr_gfp. | ||
===7.14=== | ===7.14=== | ||
| + | Do the ligation of PBAD_AfsA, Pc_ArpA, Pqrr_gfp. | ||
===7.15=== | ===7.15=== | ||
| + | Do the ligation of PBAD_AfsA, Pc_ArpA, Pqrr_gfp. | ||
===7.16=== | ===7.16=== | ||
| + | Do the ligation of PBAD_AfsA, Pc_ArpA, Pqrr_gfp. | ||
===7.17=== | ===7.17=== | ||
| + | Do the ligation of PBAD_AfsA, Pc_ArpA, Pqrr_gfp. | ||
===7.18=== | ===7.18=== | ||
| + | Do the ligation of TPP_gfp, as well as the ligation of PBAD_AfsA, Pc_ArpA, Pqrr_gfp. | ||
===7.19=== | ===7.19=== | ||
| + | Do the extraction of whole genome of Streptomyces and extract AfsA and ArpA from it, keep on doing ligation of PBAD_AfsA, Pc_ArpA, Pqrr_gfp. | ||
===7.20=== | ===7.20=== | ||
| + | Do the ligation of PBAD_TPP_gfp, as well as the ligation of PBAD_AfsA, Pc_ArpA, Pqrr_gfp. | ||
===7.21=== | ===7.21=== | ||
| + | Do the ligation of PBAD_AfsA, Pc_ArpA, Pqrr_gfp, and send plasmid of PBAD_TPP_gfp for sequencing. | ||
===7.22=== | ===7.22=== | ||
| + | Do the ligation of PBAD_AfsA, Pc_ArpA, Pqrr_gfp, and send plasmid of PBAD_TPP_gfp for sequencing. | ||
===7.23=== | ===7.23=== | ||
| + | Do the ligation of PBAD_AfsA, Pc_ArpA, Pqrr_gfp, and send plasmid of PBAD_TPP_gfp for sequencing. | ||
===7.24=== | ===7.24=== | ||
| + | Do the ligation of PBAD_AfsA, Pc_ArpA, Pqrr_gfp, and send plasmid of PBAD_TPP_gfp for sequencing. | ||
===7.25=== | ===7.25=== | ||
| + | Do the ligation of PBAD_AfsA, Pc_ArpA, Pqrr_gfp, and send plasmid of PBAD_TPP_gfp for sequencing. | ||
===7.26=== | ===7.26=== | ||
| + | Do the ligation of PBAD_AfsA, Pc_ArpA, Pqrr_gfp, and send plasmid of PBAD_TPP_gfp for sequencing. | ||
===7.27=== | ===7.27=== | ||
| + | Do the ligation of PBAD_AfsA, Pc_ArpA, Pqrr_gfp, and send plasmid of PBAD_TPP_gfp for sequencing. | ||
===7.28=== | ===7.28=== | ||
| + | The sequencing of PBAD_TPP_gfp is right, do the transformation of this part to DH5α, as well as cultivating the old E.coli containing this part in Lysogeny broth (containing ampicillin). | ||
===7.29=== | ===7.29=== | ||
| + | Cultivate the newly transformational E.coli in Lysogeny broth (containing ampicillin) and induce it with different density of TPP and under different conditions. | ||
===7.30=== | ===7.30=== | ||
| + | Cultivate the newly transformational E.coli in Lysogeny broth (containing ampicillin) and induce it with different density of TPP and under different conditions. | ||
===7.31=== | ===7.31=== | ||
| + | Cultivate the newly transformational E.coli in Lysogeny broth (containing ampicillin) and induce it with different density of TPP and under different conditions. | ||
==August== | ==August== | ||
| Line 247: | Line 286: | ||
[<html><a href="#top">TOP</a></html>] | [<html><a href="#top">TOP</a></html>] | ||
===8.1=== | ===8.1=== | ||
| + | Cultivate the newly transformational E.coli in Lysogeny broth (containing ampicillin) and induce it with different density of TPP and under different conditions. | ||
===8.2=== | ===8.2=== | ||
| + | Cultivate the newly transformational E.coli in Lysogeny broth (containing ampicillin) and induce it with different density of TPP and under different conditions. | ||
===8.3=== | ===8.3=== | ||
| + | Cultivate the newly transformational E.coli in Lysogeny broth (containing ampicillin) and induce it with different density of TPP and under different conditions. | ||
===8.4=== | ===8.4=== | ||
| + | Cultivate the newly transformational E.coli in Lysogeny broth (containing ampicillin) and induce it with different density of TPP and under different conditions. | ||
===8.5=== | ===8.5=== | ||
| + | Cultivate the newly transformational E.coli in Lysogeny broth (containing ampicillin) and induce it with different density of TPP and under different conditions. | ||
===8.6=== | ===8.6=== | ||
| + | Cultivate the newly transformational E.coli in Lysogeny broth (containing ampicillin) and induce it with different density of TPP and under different conditions. | ||
===8.7=== | ===8.7=== | ||
| + | Do the ligation of PBAD_TPP_TPP_gfp as well as keeping on doing the induction of PBAD_TPP_gfp. | ||
| + | Do the photoetching of the fourth chip and mould PDMS. | ||
===8.8=== | ===8.8=== | ||
| + | Do the ligation of PBAD_TPP_TPP_gfp as well as keeping on doing the induction of PBAD_TPP_gfp. | ||
===8.9=== | ===8.9=== | ||
| + | Do the ligation of PBAD_TPP_TPP_gfp as well as keeping on doing the induction of PBAD_TPP_gfp. | ||
===8.10=== | ===8.10=== | ||
| + | Do the ligation of PBAD_TPP_TPP_gfp as well as keeping on doing the induction of PBAD_TPP_gfp. | ||
===8.11=== | ===8.11=== | ||
| + | Do the ligation of PBAD_TPP_TPP_gfp as well as keeping on doing the induction of PBAD_TPP_gfp. | ||
===8.12=== | ===8.12=== | ||
| + | Do the ligation of PBAD_TPP_TPP_gfp as well as keeping on doing the induction of PBAD_TPP_gfp. | ||
===8.15=== | ===8.15=== | ||
| + | Do the ligation of PBAD_TPP_TPP_gfp as well as keeping on doing the induction of PBAD_TPP_gfp. | ||
===8.16=== | ===8.16=== | ||
| + | Do the ligation of PBAD_TPP_TPP_gfp as well as keeping on doing the induction of PBAD_TPP_gfp. | ||
===8.17=== | ===8.17=== | ||
| + | Do the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2), first, do the ligation of Plas_Ptsg(2’)_gfp and Prhl_Sgrs(2), respectively. Then do the ligation of the two parts together. | ||
===8.18=== | ===8.18=== | ||
| + | Do the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2), first, do the ligation of Plas_Ptsg(2’)_gfp and Prhl_Sgrs(2), respectively. Then do the ligation of the two parts together. | ||
===8.19=== | ===8.19=== | ||
| + | Do the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2), first, do the ligation of Plas_Ptsg(2’)_gfp and Prhl_Sgrs(2), respectively. Then do the ligation of the two parts together. | ||
===8.20=== | ===8.20=== | ||
| + | Do the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2), first, do the ligation of Plas_Ptsg(2’)_gfp and Prhl_Sgrs(2), respectively. Then do the ligation of the two parts together. | ||
===8.21=== | ===8.21=== | ||
| + | Do the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2), first, do the ligation of Plas_Ptsg(2’)_gfp and Prhl_Sgrs(2), respectively. Then do the ligation of the two parts together. | ||
===8.22=== | ===8.22=== | ||
| + | Do the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2), first, do the ligation of Plas_Ptsg(2’)_gfp and Prhl_Sgrs(2), respectively. Then do the ligation of the two parts together. | ||
===8.23=== | ===8.23=== | ||
| + | Do the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2), first, do the ligation of Plas_Ptsg(2’)_gfp and Prhl_Sgrs(2), respectively. Then do the ligation of the two parts together. | ||
===8.24=== | ===8.24=== | ||
| + | Do the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2), first, do the ligation of Plas_Ptsg(2’)_gfp and Prhl_Sgrs(2), respectively. Then do the ligation of the two parts together. | ||
===8.25=== | ===8.25=== | ||
| + | Do the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2), first, do the ligation of Plas_Ptsg(2’)_gfp and Prhl_Sgrs(2), respectively. Then do the ligation of the two parts together. | ||
===8.26=== | ===8.26=== | ||
| + | Do the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2), first, do the ligation of Plas_Ptsg(2’)_gfp and Prhl_Sgrs(2), respectively. Then do the ligation of the two parts together. | ||
===8.27=== | ===8.27=== | ||
| + | Do the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2), first, do the ligation of Plas_Ptsg(2’)_gfp and Prhl_Sgrs(2), respectively. Then do the ligation of the two parts together. | ||
===8.28=== | ===8.28=== | ||
| + | Do the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2), first, do the ligation of Plas_Ptsg(2’)_gfp and Prhl_Sgrs(2), respectively. Then do the ligation of the two parts together. | ||
===8.29=== | ===8.29=== | ||
| + | Do the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2), first, do the ligation of Plas_Ptsg(2’)_gfp and Prhl_Sgrs(2), respectively. Then do the ligation of the two parts together. | ||
===8.30=== | ===8.30=== | ||
| + | Do the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2), first, do the ligation of Plas_Ptsg(2’)_gfp and Prhl_Sgrs(2), respectively. Then do the ligation of the two parts together. | ||
===8.31=== | ===8.31=== | ||
| Line 358: | Line 426: | ||
[<html><a href="#top">TOP</a></html>] | [<html><a href="#top">TOP</a></html>] | ||
===9.1=== | ===9.1=== | ||
| + | Do the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2), first, do the ligation of Plas_Ptsg(2’)_gfp and Prhl_Sgrs(2), respectively. Then do the ligation of the two parts together. | ||
===9.2=== | ===9.2=== | ||
| + | Do the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2), first, do the ligation of Plas_Ptsg(2’)_gfp and Prhl_Sgrs(2), respectively. Then do the ligation of the two parts together. | ||
===9.3=== | ===9.3=== | ||
| + | Do the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2), first, do the ligation of Plas_Ptsg(2’)_gfp and Prhl_Sgrs(2), respectively. Then do the ligation of the two parts together. | ||
===9.4=== | ===9.4=== | ||
| + | Do the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2), first, do the ligation of Plas_Ptsg(2’)_gfp and Prhl_Sgrs(2), respectively. Then do the ligation of the two parts together. | ||
===9.5=== | ===9.5=== | ||
| + | Do the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2), first, do the ligation of Plas_Ptsg(2’)_gfp and Prhl_Sgrs(2), respectively. Then do the ligation of the two parts together. | ||
===9.6=== | ===9.6=== | ||
| + | Do the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2), first, do the ligation of Plas_Ptsg(2’)_gfp and Prhl_Sgrs(2), respectively. Then do the ligation of the two parts together. | ||
===9.7=== | ===9.7=== | ||
| + | Do the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2), first, do the ligation of Plas_Ptsg(2’)_gfp and Prhl_Sgrs(2), respectively. Then do the ligation of the two parts together. | ||
===9.8=== | ===9.8=== | ||
| + | Do the ligation of PT7_lasI and PT7_RBS_rhlI, while doing the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2). | ||
===9.9=== | ===9.9=== | ||
| + | Do the ligation of PT7_lasI and PT7_RBS_rhlI, while doing the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2). | ||
===9.10=== | ===9.10=== | ||
| + | Do the ligation of PT7_lasI and PT7_RBS_rhlI, while doing the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2). | ||
===9.11=== | ===9.11=== | ||
| + | Do the ligation of PT7_lasI and PT7_RBS_rhlI, while doing the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2). | ||
===9.12=== | ===9.12=== | ||
| + | Do the ligation of PT7_lasI and PT7_RBS_rhlI, while doing the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2). | ||
===9.13=== | ===9.13=== | ||
| + | Do the ligation of PT7_lasI and PT7_RBS_rhlI, while doing the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2). | ||
===9.14=== | ===9.14=== | ||
| + | Do the ligation of PT7_lasI and PT7_RBS_rhlI, while doing the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2). | ||
===9.15=== | ===9.15=== | ||
| + | Do the ligation of PT7_lasI and PT7_RBS_rhlI, while doing the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2). | ||
===9.16=== | ===9.16=== | ||
| + | Do the ligation of PT7_lasI and PT7_RBS_rhlI, while doing the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2). | ||
===9.17=== | ===9.17=== | ||
| + | Do the ligation of PT7_lasI and PT7_RBS_rhlI, while doing the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2). | ||
===9.18=== | ===9.18=== | ||
| + | Do the ligation of PT7_lasI and PT7_RBS_rhlI, while doing the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2). | ||
===9.19=== | ===9.19=== | ||
| + | Do the ligation of PT7_lasI and PT7_RBS_rhlI, while doing the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2). | ||
===9.20=== | ===9.20=== | ||
| + | Try to improve the microfluidic device and test the performance of the two-layered chips, making sure that: | ||
| + | 1、The cells grow well in the chambers. | ||
| + | 2、Different cells (RFP and GFP) are divided physically. | ||
| + | 3、The chemical wires can transfer through the membrane freely. | ||
| + | 4、The membrane and the PDMS can stick for more than 10 hours, allowing no leakage. | ||
| + | |||
| + | Of course, this part of work includes moulding chips and making two-layered chips again and again. | ||
===9.21=== | ===9.21=== | ||
| + | Try to improve the microfluidic device and test the performance of the two-layered chips, making sure that: | ||
| + | 1、The cells grow well in the chambers. | ||
| + | 2、Different cells (RFP and GFP) are divided physically. | ||
| + | 3、The chemical wires can transfer through the membrane freely. | ||
| + | 4、The membrane and the PDMS can stick for more than 10 hours, allowing no leakage. | ||
| + | |||
| + | Of course, this part of work includes moulding chips and making two-layered chips again and again. | ||
===9.22=== | ===9.22=== | ||
| + | Try to improve the microfluidic device and test the performance of the two-layered chips, making sure that: | ||
| + | 1、The cells grow well in the chambers. | ||
| + | 2、Different cells (RFP and GFP) are divided physically. | ||
| + | 3、The chemical wires can transfer through the membrane freely. | ||
| + | 4、The membrane and the PDMS can stick for more than 10 hours, allowing no leakage. | ||
| + | |||
| + | Of course, this part of work includes moulding chips and making two-layered chips again and again. | ||
===9.23=== | ===9.23=== | ||
| + | Try to improve the microfluidic device and test the performance of the two-layered chips, making sure that: | ||
| + | 1、The cells grow well in the chambers. | ||
| + | 2、Different cells (RFP and GFP) are divided physically. | ||
| + | 3、The chemical wires can transfer through the membrane freely. | ||
| + | 4、The membrane and the PDMS can stick for more than 10 hours, allowing no leakage. | ||
| + | |||
| + | Of course, this part of work includes moulding chips and making two-layered chips again and again. | ||
===9.24=== | ===9.24=== | ||
| + | Try to improve the microfluidic device and test the performance of the two-layered chips, making sure that: | ||
| + | 1、The cells grow well in the chambers. | ||
| + | 2、Different cells (RFP and GFP) are divided physically. | ||
| + | 3、The chemical wires can transfer through the membrane freely. | ||
| + | 4、The membrane and the PDMS can stick for more than 10 hours, allowing no leakage. | ||
| + | |||
| + | Of course, this part of work includes moulding chips and making two-layered chips again and again. | ||
===9.25=== | ===9.25=== | ||
| + | Try to improve the microfluidic device and test the performance of the two-layered chips, making sure that: | ||
| + | 1、The cells grow well in the chambers. | ||
| + | 2、Different cells (RFP and GFP) are divided physically. | ||
| + | 3、The chemical wires can transfer through the membrane freely. | ||
| + | 4、The membrane and the PDMS can stick for more than 10 hours, allowing no leakage. | ||
| + | |||
| + | Of course, this part of work includes moulding chips and making two-layered chips again and again. | ||
===9.26=== | ===9.26=== | ||
| + | Try to improve the microfluidic device and test the performance of the two-layered chips, making sure that: | ||
| + | 1、The cells grow well in the chambers. | ||
| + | 2、Different cells (RFP and GFP) are divided physically. | ||
| + | 3、The chemical wires can transfer through the membrane freely. | ||
| + | 4、The membrane and the PDMS can stick for more than 10 hours, allowing no leakage. | ||
| + | |||
| + | Of course, this part of work includes moulding chips and making two-layered chips again and again. | ||
===9.27=== | ===9.27=== | ||
| + | Try to improve the microfluidic device and test the performance of the two-layered chips, making sure that: | ||
| + | 1、The cells grow well in the chambers. | ||
| + | 2、Different cells (RFP and GFP) are divided physically. | ||
| + | 3、The chemical wires can transfer through the membrane freely. | ||
| + | 4、The membrane and the PDMS can stick for more than 10 hours, allowing no leakage. | ||
| + | |||
| + | Of course, this part of work includes moulding chips and making two-layered chips again and again. | ||
===9.28=== | ===9.28=== | ||
| + | Try to improve the microfluidic device and test the performance of the two-layered chips, making sure that: | ||
| + | 1、The cells grow well in the chambers. | ||
| + | 2、Different cells (RFP and GFP) are divided physically. | ||
| + | 3、The chemical wires can transfer through the membrane freely. | ||
| + | 4、The membrane and the PDMS can stick for more than 10 hours, allowing no leakage. | ||
| + | |||
| + | Of course, this part of work includes moulding chips and making two-layered chips again and again. | ||
===9.29=== | ===9.29=== | ||
| + | Try to improve the microfluidic device and test the performance of the two-layered chips, making sure that: | ||
| + | 1、The cells grow well in the chambers. | ||
| + | 2、Different cells (RFP and GFP) are divided physically. | ||
| + | 3、The chemical wires can transfer through the membrane freely. | ||
| + | 4、The membrane and the PDMS can stick for more than 10 hours, allowing no leakage. | ||
| + | |||
| + | Of course, this part of work includes moulding chips and making two-layered chips again and again. | ||
===9.30=== | ===9.30=== | ||
| + | Try to improve the microfluidic device and test the performance of the two-layered chips, making sure that: | ||
| + | 1、The cells grow well in the chambers. | ||
| + | 2、Different cells (RFP and GFP) are divided physically. | ||
| + | 3、The chemical wires can transfer through the membrane freely. | ||
| + | 4、The membrane and the PDMS can stick for more than 10 hours, allowing no leakage. | ||
| + | |||
| + | Of course, this part of work includes moulding chips and making two-layered chips again and again. | ||
==October== | ==October== | ||
| Line 434: | Line 598: | ||
|style="text-align:center"|- | |style="text-align:center"|- | ||
|style="text-align:center"|[[Team:Peking_S/lab/notebook/xjy#10.1|1]] | |style="text-align:center"|[[Team:Peking_S/lab/notebook/xjy#10.1|1]] | ||
| - | |style="text-align:center"|2 | + | |style="text-align:center"| [[Team:Peking_S/lab/notebook/xjy#10.2|2]] |
|style="text-align:center"|[[Team:Peking_S/lab/notebook/xjy#10.3|3]] | |style="text-align:center"|[[Team:Peking_S/lab/notebook/xjy#10.3|3]] | ||
|- | |- | ||
|style="text-align:center"| [[Team:Peking_S/lab/notebook/xjy#10.4|4]] | |style="text-align:center"| [[Team:Peking_S/lab/notebook/xjy#10.4|4]] | ||
|style="text-align:center"|[[Team:Peking_S/lab/notebook/xjy#10.5|5]] | |style="text-align:center"|[[Team:Peking_S/lab/notebook/xjy#10.5|5]] | ||
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| - | |style="text-align:center"| | + | |style="text-align:center"|- |
| - | |style="text-align:center"| | + | |style="text-align:center"| - |
| - | |style="text-align:center"| | + | |style="text-align:center"| - |
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|- | |- | ||
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===10.1=== | ===10.1=== | ||
| + | Write wiki of my previous work and wait for the A, B cells | ||
| + | |||
| + | ===10.2=== | ||
| + | Write wiki of my previous work and wait for the A, B cells | ||
===10.3=== | ===10.3=== | ||
| + | Write wiki of my previous work and begin to test the function of A,B cells in microfluidic system at night. | ||
===10.4=== | ===10.4=== | ||
| + | Test the function of A,B cells in microfluidic system, and take photos for demonstration of the function of A cells. | ||
===10.5=== | ===10.5=== | ||
| + | Test the funciton of A,B cells in microfluidic system, and take photos for demonstration of the function of B cells. | ||
| + | Regulate the ratio and the concentrations of the two inducers (protein generated by CinI and salicylate) and take photos for demonstration of the control over A,B cells. | ||
| + | |||
| + | Do the photoetching of the new mask. | ||
| + | |||
| + | ===10.6=== | ||
| + | Take equisite photos of cells grown in a chamber in microfluidic system and add some new content on wiki. | ||
===10.7=== | ===10.7=== | ||
Latest revision as of 20:54, 5 October 2011
Template:Https://2011.igem.org/Team:Peking S/bannerhidden
Jingyi Xi's Notebook
Summary
Mainly in charge of microfluidic device, I created a bilayer microfluidic system for our project, which provides a suitable space for different cells to be spatially separated apart and population controlled, as well as allowing chemical wires to transfer freely from each side to the other. What’s more, I also take part in molecular cloning of TPP regulator, characterization of chemical wire toolbox, and modeling work of XOR gate.
Contents
June
| Mon | Tue | Wed | Thu | Fri | Sat | Sun |
| - | - | - | - | - | 25 | 26 |
| 27 | 28 | 29 | 30 | - | - | - |
| - | - | - | - | - | - | - |
[TOP]
6.25
Design and draw blueprint of the first chip.
6.26
Do the photoetching of the first chip and mould the PDMS.
6.27
Make chips and test under microscope. Pick the relatively suitable size of chamber.
6.28
Design and draw blueprint of the second chip.
6.29
Do the photoetching of the second chip and mould the PDMS.
6.30
Make chips and test under microscope.
July
| Mon | Tue | Wed | Thu | Fri | Sat | Sun |
| - | - | - | - | 1 | 2 | 3 |
| 4 | 5 | 6 | 7 | 8 | 9 | 10 |
| 11 | 12 | 13 | 14 | 15 | 16 | 17 |
| 18 | 19 | 20 | 21 | 22 | 23 | 24 |
| 25 | 26 | 27 | 28 | 29 | 30 | 31 |
| - | - | - | - | - | - | - |
[TOP]
7.1
Change the flow velocity and test the chip under microscope, take some photos of the cells in the chamber, mould new chips.
7.2
Make chips and test under microscope with slowed flow velocity, shoot a movie of the growing population in a chamber,and mould new chips.
7.3
Test under microscope with slowed flow velocity and add a heater (37℃) to the system, the chip works well. Mould new chips.
7.4
Make chips and discussion the new chip.
7.5
Design and draw blueprint of the third chip,which has wider channels and two kinds of chamber sizes.
7.6
Do the photoetching of the third chip and mould the PDMS.
7.7
Try the two-layered chips with a membrane beside each side of chips.
7.8
Try to use photoetching machine to align the markers on the chips.
7.9
Try to use common microscope to align the markers on the chips, by hands, successfully. Take photos of both sides (RFP and GFP) under fluorescence microscope.
7.10
Design and draw blueprint of the fourth chip,with longer channels.
7.11
Do the ligation of PBAD_AfsA, Pc_ArpA, Pqrr_gfp.
7.12
Do the ligation of PBAD_AfsA, Pc_ArpA, Pqrr_gfp.
7.13
Do the ligation of PBAD_AfsA, Pc_ArpA, Pqrr_gfp.
7.14
Do the ligation of PBAD_AfsA, Pc_ArpA, Pqrr_gfp.
7.15
Do the ligation of PBAD_AfsA, Pc_ArpA, Pqrr_gfp.
7.16
Do the ligation of PBAD_AfsA, Pc_ArpA, Pqrr_gfp.
7.17
Do the ligation of PBAD_AfsA, Pc_ArpA, Pqrr_gfp.
7.18
Do the ligation of TPP_gfp, as well as the ligation of PBAD_AfsA, Pc_ArpA, Pqrr_gfp.
7.19
Do the extraction of whole genome of Streptomyces and extract AfsA and ArpA from it, keep on doing ligation of PBAD_AfsA, Pc_ArpA, Pqrr_gfp.
7.20
Do the ligation of PBAD_TPP_gfp, as well as the ligation of PBAD_AfsA, Pc_ArpA, Pqrr_gfp.
7.21
Do the ligation of PBAD_AfsA, Pc_ArpA, Pqrr_gfp, and send plasmid of PBAD_TPP_gfp for sequencing.
7.22
Do the ligation of PBAD_AfsA, Pc_ArpA, Pqrr_gfp, and send plasmid of PBAD_TPP_gfp for sequencing.
7.23
Do the ligation of PBAD_AfsA, Pc_ArpA, Pqrr_gfp, and send plasmid of PBAD_TPP_gfp for sequencing.
7.24
Do the ligation of PBAD_AfsA, Pc_ArpA, Pqrr_gfp, and send plasmid of PBAD_TPP_gfp for sequencing.
7.25
Do the ligation of PBAD_AfsA, Pc_ArpA, Pqrr_gfp, and send plasmid of PBAD_TPP_gfp for sequencing.
7.26
Do the ligation of PBAD_AfsA, Pc_ArpA, Pqrr_gfp, and send plasmid of PBAD_TPP_gfp for sequencing.
7.27
Do the ligation of PBAD_AfsA, Pc_ArpA, Pqrr_gfp, and send plasmid of PBAD_TPP_gfp for sequencing.
7.28
The sequencing of PBAD_TPP_gfp is right, do the transformation of this part to DH5α, as well as cultivating the old E.coli containing this part in Lysogeny broth (containing ampicillin).
7.29
Cultivate the newly transformational E.coli in Lysogeny broth (containing ampicillin) and induce it with different density of TPP and under different conditions.
7.30
Cultivate the newly transformational E.coli in Lysogeny broth (containing ampicillin) and induce it with different density of TPP and under different conditions.
7.31
Cultivate the newly transformational E.coli in Lysogeny broth (containing ampicillin) and induce it with different density of TPP and under different conditions.
August
| Mon | Tue | Wed | Thu | Fri | Sat | Sun |
| 1 | 2 | 3 | 4 | 5 | 6 | 7 |
| 8 | 9 | 10 | 11 | 12 | 13 | 14 |
| 15 | 16 | 17 | 18 | 19 | 20 | 21 |
| 22 | 23 | 24 | 25 | 26 | 27 | 28 |
| 29 | 30 | 31 | - | - | - | - |
[TOP]
8.1
Cultivate the newly transformational E.coli in Lysogeny broth (containing ampicillin) and induce it with different density of TPP and under different conditions.
8.2
Cultivate the newly transformational E.coli in Lysogeny broth (containing ampicillin) and induce it with different density of TPP and under different conditions.
8.3
Cultivate the newly transformational E.coli in Lysogeny broth (containing ampicillin) and induce it with different density of TPP and under different conditions.
8.4
Cultivate the newly transformational E.coli in Lysogeny broth (containing ampicillin) and induce it with different density of TPP and under different conditions.
8.5
Cultivate the newly transformational E.coli in Lysogeny broth (containing ampicillin) and induce it with different density of TPP and under different conditions.
8.6
Cultivate the newly transformational E.coli in Lysogeny broth (containing ampicillin) and induce it with different density of TPP and under different conditions.
8.7
Do the ligation of PBAD_TPP_TPP_gfp as well as keeping on doing the induction of PBAD_TPP_gfp. Do the photoetching of the fourth chip and mould PDMS.
8.8
Do the ligation of PBAD_TPP_TPP_gfp as well as keeping on doing the induction of PBAD_TPP_gfp.
8.9
Do the ligation of PBAD_TPP_TPP_gfp as well as keeping on doing the induction of PBAD_TPP_gfp.
8.10
Do the ligation of PBAD_TPP_TPP_gfp as well as keeping on doing the induction of PBAD_TPP_gfp.
8.11
Do the ligation of PBAD_TPP_TPP_gfp as well as keeping on doing the induction of PBAD_TPP_gfp.
8.12
Do the ligation of PBAD_TPP_TPP_gfp as well as keeping on doing the induction of PBAD_TPP_gfp.
8.15
Do the ligation of PBAD_TPP_TPP_gfp as well as keeping on doing the induction of PBAD_TPP_gfp.
8.16
Do the ligation of PBAD_TPP_TPP_gfp as well as keeping on doing the induction of PBAD_TPP_gfp.
8.17
Do the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2), first, do the ligation of Plas_Ptsg(2’)_gfp and Prhl_Sgrs(2), respectively. Then do the ligation of the two parts together.
8.18
Do the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2), first, do the ligation of Plas_Ptsg(2’)_gfp and Prhl_Sgrs(2), respectively. Then do the ligation of the two parts together.
8.19
Do the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2), first, do the ligation of Plas_Ptsg(2’)_gfp and Prhl_Sgrs(2), respectively. Then do the ligation of the two parts together.
8.20
Do the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2), first, do the ligation of Plas_Ptsg(2’)_gfp and Prhl_Sgrs(2), respectively. Then do the ligation of the two parts together.
8.21
Do the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2), first, do the ligation of Plas_Ptsg(2’)_gfp and Prhl_Sgrs(2), respectively. Then do the ligation of the two parts together.
8.22
Do the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2), first, do the ligation of Plas_Ptsg(2’)_gfp and Prhl_Sgrs(2), respectively. Then do the ligation of the two parts together.
8.23
Do the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2), first, do the ligation of Plas_Ptsg(2’)_gfp and Prhl_Sgrs(2), respectively. Then do the ligation of the two parts together.
8.24
Do the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2), first, do the ligation of Plas_Ptsg(2’)_gfp and Prhl_Sgrs(2), respectively. Then do the ligation of the two parts together.
8.25
Do the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2), first, do the ligation of Plas_Ptsg(2’)_gfp and Prhl_Sgrs(2), respectively. Then do the ligation of the two parts together.
8.26
Do the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2), first, do the ligation of Plas_Ptsg(2’)_gfp and Prhl_Sgrs(2), respectively. Then do the ligation of the two parts together.
8.27
Do the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2), first, do the ligation of Plas_Ptsg(2’)_gfp and Prhl_Sgrs(2), respectively. Then do the ligation of the two parts together.
8.28
Do the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2), first, do the ligation of Plas_Ptsg(2’)_gfp and Prhl_Sgrs(2), respectively. Then do the ligation of the two parts together.
8.29
Do the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2), first, do the ligation of Plas_Ptsg(2’)_gfp and Prhl_Sgrs(2), respectively. Then do the ligation of the two parts together.
8.30
Do the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2), first, do the ligation of Plas_Ptsg(2’)_gfp and Prhl_Sgrs(2), respectively. Then do the ligation of the two parts together.
8.31
September
| Mon | Tue | Wed | Thu | Fri | Sat | Sun |
| - | - | 1 | 2 | 3 | 4 | |
| 5 | 6 | 7 | 8 | 9 | 10 | 11 |
| 12 | 13 | 14 | 15 | 16 | 17 | 18 |
| 19 | 20 | 21 | 22 | 23 | 24 | 25 |
| 26 | 27 | 28 | 29 | 30 | - | - |
[TOP]
9.1
Do the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2), first, do the ligation of Plas_Ptsg(2’)_gfp and Prhl_Sgrs(2), respectively. Then do the ligation of the two parts together.
9.2
Do the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2), first, do the ligation of Plas_Ptsg(2’)_gfp and Prhl_Sgrs(2), respectively. Then do the ligation of the two parts together.
9.3
Do the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2), first, do the ligation of Plas_Ptsg(2’)_gfp and Prhl_Sgrs(2), respectively. Then do the ligation of the two parts together.
9.4
Do the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2), first, do the ligation of Plas_Ptsg(2’)_gfp and Prhl_Sgrs(2), respectively. Then do the ligation of the two parts together.
9.5
Do the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2), first, do the ligation of Plas_Ptsg(2’)_gfp and Prhl_Sgrs(2), respectively. Then do the ligation of the two parts together.
9.6
Do the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2), first, do the ligation of Plas_Ptsg(2’)_gfp and Prhl_Sgrs(2), respectively. Then do the ligation of the two parts together.
9.7
Do the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2), first, do the ligation of Plas_Ptsg(2’)_gfp and Prhl_Sgrs(2), respectively. Then do the ligation of the two parts together.
9.8
Do the ligation of PT7_lasI and PT7_RBS_rhlI, while doing the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2).
9.9
Do the ligation of PT7_lasI and PT7_RBS_rhlI, while doing the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2).
9.10
Do the ligation of PT7_lasI and PT7_RBS_rhlI, while doing the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2).
9.11
Do the ligation of PT7_lasI and PT7_RBS_rhlI, while doing the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2).
9.12
Do the ligation of PT7_lasI and PT7_RBS_rhlI, while doing the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2).
9.13
Do the ligation of PT7_lasI and PT7_RBS_rhlI, while doing the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2).
9.14
Do the ligation of PT7_lasI and PT7_RBS_rhlI, while doing the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2).
9.15
Do the ligation of PT7_lasI and PT7_RBS_rhlI, while doing the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2).
9.16
Do the ligation of PT7_lasI and PT7_RBS_rhlI, while doing the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2).
9.17
Do the ligation of PT7_lasI and PT7_RBS_rhlI, while doing the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2).
9.18
Do the ligation of PT7_lasI and PT7_RBS_rhlI, while doing the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2).
9.19
Do the ligation of PT7_lasI and PT7_RBS_rhlI, while doing the ligation of Plas_Ptsg(2’) _gfp_Prhl_Sgrs(2).
9.20
Try to improve the microfluidic device and test the performance of the two-layered chips, making sure that: 1、The cells grow well in the chambers. 2、Different cells (RFP and GFP) are divided physically. 3、The chemical wires can transfer through the membrane freely. 4、The membrane and the PDMS can stick for more than 10 hours, allowing no leakage.
Of course, this part of work includes moulding chips and making two-layered chips again and again.
9.21
Try to improve the microfluidic device and test the performance of the two-layered chips, making sure that: 1、The cells grow well in the chambers. 2、Different cells (RFP and GFP) are divided physically. 3、The chemical wires can transfer through the membrane freely. 4、The membrane and the PDMS can stick for more than 10 hours, allowing no leakage.
Of course, this part of work includes moulding chips and making two-layered chips again and again.
9.22
Try to improve the microfluidic device and test the performance of the two-layered chips, making sure that: 1、The cells grow well in the chambers. 2、Different cells (RFP and GFP) are divided physically. 3、The chemical wires can transfer through the membrane freely. 4、The membrane and the PDMS can stick for more than 10 hours, allowing no leakage.
Of course, this part of work includes moulding chips and making two-layered chips again and again.
9.23
Try to improve the microfluidic device and test the performance of the two-layered chips, making sure that: 1、The cells grow well in the chambers. 2、Different cells (RFP and GFP) are divided physically. 3、The chemical wires can transfer through the membrane freely. 4、The membrane and the PDMS can stick for more than 10 hours, allowing no leakage.
Of course, this part of work includes moulding chips and making two-layered chips again and again.
9.24
Try to improve the microfluidic device and test the performance of the two-layered chips, making sure that: 1、The cells grow well in the chambers. 2、Different cells (RFP and GFP) are divided physically. 3、The chemical wires can transfer through the membrane freely. 4、The membrane and the PDMS can stick for more than 10 hours, allowing no leakage.
Of course, this part of work includes moulding chips and making two-layered chips again and again.
9.25
Try to improve the microfluidic device and test the performance of the two-layered chips, making sure that: 1、The cells grow well in the chambers. 2、Different cells (RFP and GFP) are divided physically. 3、The chemical wires can transfer through the membrane freely. 4、The membrane and the PDMS can stick for more than 10 hours, allowing no leakage.
Of course, this part of work includes moulding chips and making two-layered chips again and again.
9.26
Try to improve the microfluidic device and test the performance of the two-layered chips, making sure that: 1、The cells grow well in the chambers. 2、Different cells (RFP and GFP) are divided physically. 3、The chemical wires can transfer through the membrane freely. 4、The membrane and the PDMS can stick for more than 10 hours, allowing no leakage.
Of course, this part of work includes moulding chips and making two-layered chips again and again.
9.27
Try to improve the microfluidic device and test the performance of the two-layered chips, making sure that: 1、The cells grow well in the chambers. 2、Different cells (RFP and GFP) are divided physically. 3、The chemical wires can transfer through the membrane freely. 4、The membrane and the PDMS can stick for more than 10 hours, allowing no leakage.
Of course, this part of work includes moulding chips and making two-layered chips again and again.
9.28
Try to improve the microfluidic device and test the performance of the two-layered chips, making sure that: 1、The cells grow well in the chambers. 2、Different cells (RFP and GFP) are divided physically. 3、The chemical wires can transfer through the membrane freely. 4、The membrane and the PDMS can stick for more than 10 hours, allowing no leakage.
Of course, this part of work includes moulding chips and making two-layered chips again and again.
9.29
Try to improve the microfluidic device and test the performance of the two-layered chips, making sure that: 1、The cells grow well in the chambers. 2、Different cells (RFP and GFP) are divided physically. 3、The chemical wires can transfer through the membrane freely. 4、The membrane and the PDMS can stick for more than 10 hours, allowing no leakage.
Of course, this part of work includes moulding chips and making two-layered chips again and again.
9.30
Try to improve the microfluidic device and test the performance of the two-layered chips, making sure that: 1、The cells grow well in the chambers. 2、Different cells (RFP and GFP) are divided physically. 3、The chemical wires can transfer through the membrane freely. 4、The membrane and the PDMS can stick for more than 10 hours, allowing no leakage.
Of course, this part of work includes moulding chips and making two-layered chips again and again.
October
| Mon | Tue | Wed | Thu | Fri | Sat | Sun |
| - | - | - | - | 1 | 2 | 3 |
| 4 | 5 | - | - | - | - | - |
| - | - | - | - | - | - | - |
| - | - | - | - | - | - | - |
| - | - | - | - | - | - | - |
[TOP]
10.1
Write wiki of my previous work and wait for the A, B cells
10.2
Write wiki of my previous work and wait for the A, B cells
10.3
Write wiki of my previous work and begin to test the function of A,B cells in microfluidic system at night.
10.4
Test the function of A,B cells in microfluidic system, and take photos for demonstration of the function of A cells.
10.5
Test the funciton of A,B cells in microfluidic system, and take photos for demonstration of the function of B cells. Regulate the ratio and the concentrations of the two inducers (protein generated by CinI and salicylate) and take photos for demonstration of the control over A,B cells.
Do the photoetching of the new mask.
10.6
Take equisite photos of cells grown in a chamber in microfluidic system and add some new content on wiki.
10.7
10.8
10.9
10.10
10.11
10.12
10.13
10.15
10.16-10.21
10.21-10.25
