Sudoku

Abstract Construct Experiment Notebook Result

Notebook

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 digestion

2, 3, 5, 9 plasmid (6/25 miniprep)

2 : 59.3 ng/uL SP

3 : 124.2 ng/uL XP

5 : 81.3 ng/uL SP

9 : 85.4 ng/uL SP

Gel extract

2, 3, 5, 9 plasmid (7/1 enzyme digestion)

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 digestion

2-3, 11 plasmid (7/2 miniprep)

2-3 : ES

11 (1st) : ES -> Judged failure by cut check → miniprep again

11 (2nd) : ES

7/3

Gel extract

2-3, 11 plasmid (7/2 enzyme digestion)

Label : 100703 X ゲル抽出

7/5

Ligation

2-3 (insert, 7/3 ES enzyme digestion) & 4 (vector, probably 6/28 gel extract, EX enzyme digestion) / 11 (insert; 7/3 ES enzyme digestion) & 7 (vector, probably 6/28 gel extract, EX enzyme digestion)

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)

7/6

Inoculation

234 plasmid (7/5 plate)

11-7 plasmid (7/5 plate) -> failure

Miniprep

234 plasmid (7/6 plate)

234 I -> 84.9 ng/uL

234 II -> 98.9 ng/uL

234 III -> 98.1 ng/uL

234 IV -> 99.4 ng/uL

Enzyme digestion

11 (7/1 miniprep)

11 : ES

ethanol precipitation

failure

Inoculation

7 plasmid (6/23 plate)

Enzyme digestion

1, 4 (6/28 enzyme digestion -> short), 11 plasmid (7/6 miniprep)

1 : SP

4 : EX

234 : XP

Inoculation

5 plasmid (6/23 plate)

7/7

Gel extract

1, 4, 234 ,11 plasmid (7/6 enzyme digestion)

Miniprep

5, 7 plasmid (7/6 plate)

5 -> 25.5 ng/uL

7 -> 22.3 ng/uL

Enzyme digestion

5,7 (7/7 miniprep)

5 : SP

7 : EX

Gel extract

5, 7 plasmid (7/7 enzyme digestion)

Ligation

1 (7/7 gel extract) & 234 (7/7 gel extract) / 9 (7/1 gel extract) & 234 (7/7 gel extract) / 7 (7/7 gel extract) & 11 (7/7 gel extract)

7/8

Enzyme digestion

11 (7/7 gel extract ,4.1 ng/uL), 7

11 : ES

7 : EX

Gel extract

7 (7/6 enzyme digestion), 234 (7/7 enzyme digestion) plasmid

Ligation

234(7/8 gel extract) & 9(7/1 gel extract)

234 : 10.2 ng/uL -> 4.1 uL & 9 : 10.6 ng/uL -> 5.9 uL

7/9

Transformation

9-234 plasmid (7/8 ligation)

Gel extraction

11 plasmid (7/8 enzyme digestion)

Cut check

7 (7/8 enzyme digestion)

Inoculation

1-234 plasmid (7/7 transformation), 9 plasmid (6/23)

Ligation

11 (7/1 ligation) & 7 (7/8 ligation)

11 : 6.5 ng/uL

7/10

Miniprep

1-234 (7/9 inoculation)

1-234 : 107.2 ng/uL

Transformation

9-234

Inoculation

9 (6/23 transformation)

7/11

enzyme digestion

7 (7/7 miniprep), 234 (7/6 miniprep)

7 : EX

234 : XP

Inoculation

9-234 (7/9 transformation -> failure), 11-7 (7/10 transformation), 9 (6/23 transformation), 11 (6/30 transformation)

7/12

Miniprep

11-7, 9, 11 (7/11 inoculation)

11-7 : 81.2 ng/uL

9 : 84.4 ng/uL

11 : 81.2 ng/uL

Enzyme digestion

11-7, 9 (7/12 miniprep)

11-7 : XP

9 : SP

Cut check

1-234 (7/10 107.2 ng/uL), 234 (7/6 98.1 ng/uL), 2-3 (7/2 105.0 ng/uL), 3 (6/25 124.2 ng/uL) plasmid

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)