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Team:Todai-Tokyo/Notebook/bioclock
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=='''August/September'''== | =='''August/September'''== | ||
*create plasmid including EcoRI restriction site, XbaI restriction site, placI/araC, XhoI restriction site, NcoI restriction site, double terminator, SpeI restriction site and PstI restriction site.→pUC57<BR> | *create plasmid including EcoRI restriction site, XbaI restriction site, placI/araC, XhoI restriction site, NcoI restriction site, double terminator, SpeI restriction site and PstI restriction site.→pUC57<BR> | ||
| - | -E-X-placI/araC-XhoI-NcoI-double terminator-P-<BR> | + | -E-X-placI/araC-XhoI-NcoI-double terminator-S-P-<BR> |
*cut pUC57 by XhoI and NcoI<BR> | *cut pUC57 by XhoI and NcoI<BR> | ||
→insert GFP, araC or lacI<BR> | →insert GFP, araC or lacI<BR> | ||
| - | (GFP:2006 plate1-16E,araC and lacI:2009 iGEM | + | (GFP:2006 plate1-16E,araC and lacI:2009 iGEM distribution) |
*cut plate1-23L(double terminator) and insert plate1-7L(lacI) in it<BR> | *cut plate1-23L(double terminator) and insert plate1-7L(lacI) in it<BR> | ||
*PCR of cI~cII<BR> | *PCR of cI~cII<BR> | ||
Revision as of 04:50, 19 October 2009
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Contents |
Plan
Aim: Create E.coli that show oscillatory gene expression pattern (colE1 ori vector)-placI/araC-araC+ssra tag-double terminator-placI/araC-GFP+ssra tag-double terminator- (p15A ori)-placI/araC-lacI+ssra tag-double terminator-
Methods:
1. Clone the following genes from Bacillus subtilis into a biobrick vector.
- plate1,13B
- plate1,13L
- plate2,1H
2. Make a gene network that express oscillatory pattern, using these genes.
7/7
Cloning the parts
preculture of the Biobrick parts for Miniprep
7/8
Cloning the parts
Miniprep of E.coli cells containing Biobrick parts with Promega, Wizard Plus SV Miniprep DNA Purification System
failure
7/9
Cloning the parts
Miniprep again of E.coli cells containing Biobrick parts with Promega, Wizard Plus SV Miniprep DNA Purification System
7/29
Miniprep
- P1.14L(araC)
- P1.7L(lacI)
- P1.4E(cI)
- P1.3D(ColE1)
- P1.9C(p15A)
- P1.9G(p15A)
7/30
Miniprep
- P1.14L(araC)
- P1.7L(lacI)
- P3.21D
August/September
- create plasmid including EcoRI restriction site, XbaI restriction site, placI/araC, XhoI restriction site, NcoI restriction site, double terminator, SpeI restriction site and PstI restriction site.→pUC57
-E-X-placI/araC-XhoI-NcoI-double terminator-S-P-
- cut pUC57 by XhoI and NcoI
→insert GFP, araC or lacI
(GFP:2006 plate1-16E,araC and lacI:2009 iGEM distribution)
- cut plate1-23L(double terminator) and insert plate1-7L(lacI) in it
- PCR of cI~cII
October
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