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)