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Team:UT-Tokyo/Sudoku notebook
From 2010.igem.org
(→7/6) |
(→Pre-experiment) |
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'''Enzyme digestion''' | '''Enzyme digestion''' | ||
| - | 2, 3, 5, | + | 2, 3, 5, 9 plasmid (6/25 miniprep) |
2 : 59.3 ng/uL SP | 2 : 59.3 ng/uL SP | ||
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5 : 81.3 ng/uL SP | 5 : 81.3 ng/uL SP | ||
| - | + | 9 : 85.4 ng/uL SP | |
'''Gel extract''' | '''Gel extract''' | ||
| - | 2, 3, 5, | + | 2, 3, 5, 9 plasmid (7/1 enzyme digestion) |
Label : 100701 数X (2,3,5,9) ゲル抽出 | Label : 100701 数X (2,3,5,9) ゲル抽出 | ||
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5 plasmid (6/23 plate) | 5 plasmid (6/23 plate) | ||
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==== 7/7 ==== | ==== 7/7 ==== | ||
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1, 4, 234 ,11 plasmid (7/6 enzyme digestion) | 1, 4, 234 ,11 plasmid (7/6 enzyme digestion) | ||
| + | |||
| + | |||
| + | '''Miniprep''' | ||
| + | |||
| + | 5, 7 plasmid (7/6 plate) | ||
| + | |||
| + | 5 -> 25.5 ng/uL | ||
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| + | 7 -> 22.3 ng/uL | ||
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| + | |||
| + | '''Enzyme digestion''' | ||
| + | |||
| + | 5,7 | ||
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| + | 5 : SP | ||
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| + | 7 : EX | ||
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| + | |||
| + | '''Gel extract''' | ||
| + | |||
| + | 5, 7 plasmid | ||
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| + | |||
| + | '''Ligation''' | ||
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| + | 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) | ||
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| + | |||
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| + | ==== 7/8 ==== | ||
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=== Check test === | === Check test === | ||
Revision as of 04:14, 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
5 : SP
7 : EX
Gel extract
5, 7 plasmid
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
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)
