Team:UT-Tokyo/Sudoku data
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
2, 3, 5, 7 plasmid(6/25 miniprep)
2 : 53.9 ng/uL → about 15 uL(shortage) / H / SP(2 uL)
3 : 124.2 ng/uL → 8 uL / M / XP(2 uL)
5 : 81.3 ng/uL → 12.3 uL / H / SP(2 uL)
7 : 85.4 ng/uL → 11.7 uL / H / SP(2 uL)
Gel extract
2, 3, 5, 7 plasmid(7/1 enzyme cut)
Label: 100701 数X(2,3,5,9) ゲル抽出
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)
Check test
Check test
Number | Name | Link to BioBrick | Plate coordinate | Vector | Code length |
1 | T7 promoter | plate1-6N | pSB1AK8 | 46bp | |
2 | rbs | plate1-1H | pSB1A2 | 15bp | |
3 | cre recombinase | plate1-5D | pSB1A2 | 1037bp | |
4 | double terminator | plate2-24C | pSB1AK3 | 95bp | |
5 | lox66 recombinase site | plate1-17J | pSB1A2 | 34bp | |
6 | Kan resistance (rev) | plate1-2K | pSB1A2 | 816bp | |
7 | rbs (rev) | plate1-1J | pSB1A2 | 15bp | |
8 | lox71 recombinase site |
order-made | --- | --- | 34bp |
9 | single terminator | plate1-4H | pSB1AK3 | 39bp | |
10 | constant express promoter | plate1-18A | pSB1A2 | 35bp | |
11 | Tet resistance (rev) | 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)