Team:UT-Tokyo/Sudoku notebook

From 2010.igem.org

(Difference between revisions)
(7/5)
(Pre-experiment)
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==== 7/13 ====
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'''Gel extraction'''
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11-7, 9 (7/12 enzyme digestion), 2-3-4 (7/11 enzyme digestion)
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'''Ligation'''
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9 & 2-3-4 (7/13 gel extract)/ 5 (7/7 gel extract) & 11-7 (7/13 gel extract)
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==== 7/14 ====
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'''Transformation'''
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5-11-7, 9-234 (7/13 gel extract)
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'''Inoculation'''
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5-11-7, 9-234 (7/14 transformation)
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==== 7/15 ====
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'''Miniprep'''
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5-11-7, 9-234 (7/14 inoculation)
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5-11-7 I 62.6 ng/uL
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5-11-7 II 82.6 ng/uL
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5-11-7 III 59.5 ng/uL
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9-234 I 46.4 ng/uL
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9-234 II 50.6 ng/uL
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9-234 III 41.8 ng/uL
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'''Enzyme digestion'''
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5-11-7, 9-234 (7/15 miniprep)
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==== 7/16 ====
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'''Inoculation'''
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9-234 (7/14 plate)
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'''Ligation'''
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9 & 234 (7/13 gel extract) / 1 (7/7 gel extract) & 5-11-7 (7/16 gel extract)
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'''Enzyme digestion'''
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9 (7/12 miniprep)
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9 : SP
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'''Gel extraction'''
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9-234, 5-11-7 (7/15 enzyme digestion)
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==== 7/17 ====
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'''Miniprep'''
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9-234 (7/16 inoculation)
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9-234 I 35.0 ng/uL
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9-234 II 75.4 ng/uL
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'''Enzyme digestion'''
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9-234 (7/17 miniprep)
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'''Gel extraction'''
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9 (7/16 enzyme digestion)
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'''Transformation'''
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9-234, 1-5-11-7, 9 (7/16 ligation)
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==== 7/18 ====
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'''Gel extract'''
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9-234 (7/17 enzyme digestion)
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'''Ligation'''
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9 (vector, 7/17 SP gel extract) & 234 (insert, 7/13 XP gel extract)
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9 : 10.8 ng/uL -> 5 uL & 234 : 7.1 ng/uL -> 5 uL
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'''Ligation'''
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9 (vector, 7/17 SP gel extract) & 9234 (insert, 7/18 XP gel extract) / 1 (vector, 7/7 SP gel extract) & 9234 (insert, 7/18 XP gel extract)
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9 : 10.8 ng/uL -> 5 uL & 9234 : 8.6 ng/uL -> 5 uL
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1 : 28.4 ng/uL -> 3 uL & 9234 : 8.6 ng/uL -> 7 uL
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'''Gel extract'''
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9-234 (7/17 XP enzyme digestion)
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==== 7/19 ====
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'''Transformation'''
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9-9234, 1-9234 (7/18 ligation)
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'''Miniprep'''
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1-5-11-7 (7/17 inoculation)
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1-5-11-7 I : 142.1 ng/uL
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1-5-11-7 II : 143.9 ng/uL
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1-5-11-7 III : 131.5 ng/uL
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==== 7/20 ====
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'''Sequencing'''
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9-234 II(7/17 miniprep)
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1-234 (7/10 miniprep)
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234 (7/6 miniprep)
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1-5-11-7 (7/19 miniprep)
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'''Inoculation'''
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9-9234, 1-9234 (7/19 plate)
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==== 7/21 ====
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'''Miniprep'''
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1-9234, 9-9234 (7/20 inoculation)
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1-9234 I 69.4 ng/uL
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1-9234 II 58.7 ng/uL
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1-9234 III 50.5 ng/uL
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1-9234 IV 68.8 ng/uL
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9-9234 I 100.3 ng/uL
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9-9234 II 29.4 ng/uL
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9-9234 III 76.8 ng/uL
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9-9234 IV 70.3 ng/uL
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'''Enzyme digestion'''
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9-9234 I,III (7/21 miniprep) : XP
=== Check test ===
=== Check test ===

Revision as of 07:44, 19 September 2010

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 extract

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


7/13

Gel extraction

11-7, 9 (7/12 enzyme digestion), 2-3-4 (7/11 enzyme digestion)


Ligation

9 & 2-3-4 (7/13 gel extract)/ 5 (7/7 gel extract) & 11-7 (7/13 gel extract)


7/14

Transformation

5-11-7, 9-234 (7/13 gel extract)


Inoculation

5-11-7, 9-234 (7/14 transformation)


7/15

Miniprep

5-11-7, 9-234 (7/14 inoculation)

5-11-7 I 62.6 ng/uL

5-11-7 II 82.6 ng/uL

5-11-7 III 59.5 ng/uL

9-234 I 46.4 ng/uL

9-234 II 50.6 ng/uL

9-234 III 41.8 ng/uL


Enzyme digestion

5-11-7, 9-234 (7/15 miniprep)


7/16

Inoculation

9-234 (7/14 plate)


Ligation

9 & 234 (7/13 gel extract) / 1 (7/7 gel extract) & 5-11-7 (7/16 gel extract)


Enzyme digestion

9 (7/12 miniprep)

9 : SP


Gel extraction

9-234, 5-11-7 (7/15 enzyme digestion)


7/17

Miniprep

9-234 (7/16 inoculation)

9-234 I 35.0 ng/uL

9-234 II 75.4 ng/uL


Enzyme digestion

9-234 (7/17 miniprep)


Gel extraction

9 (7/16 enzyme digestion)


Transformation

9-234, 1-5-11-7, 9 (7/16 ligation)


7/18

Gel extract

9-234 (7/17 enzyme digestion)


Ligation

9 (vector, 7/17 SP gel extract) & 234 (insert, 7/13 XP gel extract)

9 : 10.8 ng/uL -> 5 uL & 234 : 7.1 ng/uL -> 5 uL


Ligation

9 (vector, 7/17 SP gel extract) & 9234 (insert, 7/18 XP gel extract) / 1 (vector, 7/7 SP gel extract) & 9234 (insert, 7/18 XP gel extract)

9 : 10.8 ng/uL -> 5 uL & 9234 : 8.6 ng/uL -> 5 uL

1 : 28.4 ng/uL -> 3 uL & 9234 : 8.6 ng/uL -> 7 uL


Gel extract

9-234 (7/17 XP enzyme digestion)


7/19

Transformation

9-9234, 1-9234 (7/18 ligation)


Miniprep

1-5-11-7 (7/17 inoculation)

1-5-11-7 I : 142.1 ng/uL

1-5-11-7 II : 143.9 ng/uL

1-5-11-7 III : 131.5 ng/uL


7/20

Sequencing

9-234 II(7/17 miniprep)

1-234 (7/10 miniprep)

234 (7/6 miniprep)

1-5-11-7 (7/19 miniprep)


Inoculation

9-9234, 1-9234 (7/19 plate)


7/21

Miniprep

1-9234, 9-9234 (7/20 inoculation)

1-9234 I 69.4 ng/uL

1-9234 II 58.7 ng/uL

1-9234 III 50.5 ng/uL

1-9234 IV 68.8 ng/uL

9-9234 I 100.3 ng/uL

9-9234 II 29.4 ng/uL

9-9234 III 76.8 ng/uL

9-9234 IV 70.3 ng/uL


Enzyme digestion

9-9234 I,III (7/21 miniprep) : XP

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)

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