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Team:UT-Tokyo/Sudoku notebook 2
From 2010.igem.org
(New page: {{UT-Tokyo_HEADER}} <div id="main_all"> = '''Sudoku''' = [https://2010.igem.org/Team:UT-Tokyo/Sudoku_abstract Abstract] [https://2010.igem.org/Team:UT-Tokyo/Sudoku_construct Constru...) |
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=== Make construct === | === Make construct === | ||
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| + | sequel to 7/31 | ||
| + | add HiDi 20μL | ||
| + | A6 I pre, B6 I sur,C6 II pre | ||
| + | D6 II sur, E6 III pre, F6 III sur | ||
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2-3-4 plasmid (7/5 ligation) / 11-7 plasmid (7/5 ligation) | 2-3-4 plasmid (7/5 ligation) / 11-7 plasmid (7/5 ligation) | ||
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=== Check test === | === Check test === | ||
Revision as of 01:40, 19 September 2010
Sudoku
Abstract Construct Experiment Notebook Result
Notebook
Pre-experiment
AUGUST!!!!!
AUGUST!!!!!
AUGUST!!!!!
AUGUST!!!!!
AUGUST!!!!!
Make construct
8/2
Sequencing
sequel to 7/31 add HiDi 20μL A6 I pre, B6 I sur,C6 II pre D6 II sur, E6 III pre, F6 III sur
7/2
Miniprep
2-3, 11 plasmid (7/1 inoculation)
2-3 → 105 ng/uL
11 (1st) → 191.3 ng/uL
11 (2nd) → 49.5 ng/uL
Enzyme cut
2-3, 11 plasmid (7/2 miniprep)
2-3 : 9.5 uL / H / ES (1.5 uL)
11 (1st) : 5.2 uL / H / ES (1.5 uL) → Judged failure by cut check → miniprep again
11 (2nd) : 20.2 uL / H / ES (1.5 uL)
7/3
Gel extract
2-3, 11 plasmid (7/2 enzyme cut)
Label : 100703 X ゲル抽出
7/5
Ligation
2-3 (7/3 enzyme cut) & 4 / 11 (7/3 enzyme cut) & 7
2-3 : 37.5 ng/uL → 5 uL & 4 : 5.2 ng/uL → 5 uL
11 : 4.2 ng/uL → 7.5 uL & 7 : Unknown → 2.5 uL
Transformation
2-3-4 plasmid (7/5 ligation) / 11-7 plasmid (7/5 ligation)
Check test
Main construct
Research of SP6 RNApol cDNA and F plasmid
Convert lox71(rev) to lox66(rev)
Make Hin(rev)
Revise MS2 gene
Make location(rev)-rbs(rev)-location(rev)
