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Team:UT-Tokyo/Sudoku protocols
From 2010.igem.org
Sudoku
Data
Pre-experiment
Make construct
7/1
Transformation
2-3 plasmid(6/30 ligation)
<6/30 transformation → fail>
Inoculation
11 plasmid(6/30 thaw, transformation)
Enzyme cut & Gel extract
2,3,5,7 plasmid(6/25 miniprep)
2:53.9 ng/uL → about 15 uL(shortage)
3:124.2 ng/uL → 8 uL
5:81.3 ng/uL → 12.3 uL
7:85.4 ng/uL → 11.7 uL
Label: 100701 数X(2,3,5,9) ゲル抽出
Check test
Check test
| Number | Name | Link to BioBrick | Plate coordinate | Vector | Code length |
| 1 | T7 promoter |
[http://partsregistry.org/wiki/index.php?title=Part:BBa_I712074 BBa_I712074] | plate1-6N | pSB1AK8 | 46bp |
| 2 | rbs |
[http://partsregistry.org/wiki/index.php?title=Part:BBa_B0030 BBa_B0030] | plate1-1H | pSB1A2 | 15bp |
| 3 | cre recombinase |
[http://partsregistry.org/wiki/index.php?title=Part:BBa_J61047 BBa_J61047] | plate1-5D | pSB1A2 | 1037bp |
| 4 | double terminator |
[http://partsregistry.org/wiki/index.php?title=Part:BBa_B0014 BBa_B0014] | plate2-24C | pSB1AK3 | 95bp |
| 5 | lox66 recombinase site |
[http://partsregistry.org/wiki/index.php?title=Part:BBa_I718017 BBa_I718017] | plate1-17J | pSB1A2 | 34bp |
| 6 | Kan resistance (rev) |
[http://partsregistry.org/wiki/index.php?title=Part:BBa_J31002 BBa_J31002] | plate1-2K | pSB1A2 | 816bp |
| 7 | rbs (rev) |
[http://partsregistry.org/wiki/index.php?title=Part:BBa_B0014 BBa_B0014] | plate1-1J | pSB1A2 | 15bp |
| 8 | lox71 recombinase site |
order-made | --- | --- | 34bp |
| 9 | single terminator |
[http://partsregistry.org/wiki/index.php?title=Part:BBa_B1006 BBa_B1006] | plate1-4H | pSB1AK3 | 39bp |
| 10 | constant express promoter |
[http://partsregistry.org/wiki/index.php?title=Part:BBa_J23119 BBa_J23119] | plate1-18A | pSB1A2 | 35bp |
| 11 | Tet resistance (rev) |
[http://partsregistry.org/wiki/index.php?title=Part:BBa_J31006 BBa_J31006] | plate1-1N | pSB1A2 | 1191bp |
Main construct
Research of SP6 RNApol cDNA and F plasmid
Research of SP6 RNApol cDNA and F plasmid
Convert lox71(rev) to lox66(rev)
Convert lox71(rev) to lox66(rev)
Make Hin(rev)
Make Hin(rev)
Revise MS2 gene
Revise MS2 gene
Make location(rev)-rbs(rev)-location(rev)
Make location(rev)-rbs(rev)-location(rev)
