Team:UT-Tokyo/Sudoku data

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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

BBa_I712074

plate1-6N pSB1AK8 46bp
2 rbs

BBa_B0030

plate1-1H pSB1A2 15bp
3 cre recombinase

BBa_J61047

plate1-5D pSB1A2 1037bp
4 double terminator

BBa_B0014

plate2-24C pSB1AK3 95bp
5 lox66 recombinase site

BBa_I718017

plate1-17J pSB1A2 34bp
6 Kan resistance (rev)

BBa_J31002

plate1-2K pSB1A2 816bp
7 rbs (rev)

BBa_B0014

plate1-1J pSB1A2 15bp
8 lox71 recombinase site

order-made

--- --- 34bp
9 single terminator

BBa_B1006

plate1-4H pSB1AK3 39bp
10 constant express promoter

BBa_J23119

plate1-18A pSB1A2 35bp
11 Tet resistance (rev)

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)


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