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

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)