From 2013.igem.org
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Revision as of 13:19, 25 September 2013
Sep 7
Miniprep
No name
DNA | concentration[µg/mL] | 260/280 | 260/230
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RBS-lysis1-DT | 195.3 | |
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RBS-lysis3-DT | 255.5 | 1.91 | 2.04
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Ptet-pT18 | 183.6 | 1.85 | 1.48
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Pcon-RBS-tetR-DT | 134.3 | 1.94 | 2.20
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Ptet-RBS-lacZα-DT | 67.7 | 1.92 | 1.79
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Pcon-pT181 attenuator | 232.3 | 1.95 | 1.90
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pSB4K5 | 237.0 | 1.61 | 1.50
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pT181 antisense(pSB1C3) | 256.4 | 1.63 | 1.78
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apt12_1R(pSB1C3) | 230.0 | 1.98 | 2.28
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Electrophoresis
No name
Lane | Sample
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1 | 100bp ladder
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2 | Plac-Spinach-DT -5
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3 | Plac-Spinach-DT -6
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4 | Plac-Spinach-DT -7
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5 | Plac-Spinach-DT -8
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6 | Plac-Spinach-DT -9
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7 | Plac-Spinach-DT -10
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8 | 100bp ladder
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1 | 100bp ladder
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2 | Plac-Spinach-DT -11
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3 | Plac-Spinach-DT -12
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4 | Pcon-antisense -5
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5 | Pcon-antisense -6
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6 | Pcon-antisense -7
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7 | Pcon-antisense -8
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8 | 100bp ladder
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Lane | Sample
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1 | 100bp ladder
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2 | Pcon-antisense -9
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3 | Pcon-antisense -10
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4 | Pcon-antisense -11
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5 | Pcon-antisense -12
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6 | Plac-attenuator -5
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7 | Plac-attenuator -6
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8 | 100bp ladder
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1 | 100bp ladder
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2 | Plac-attenuator -7
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3 | Plac-attenuator -8
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4 | Plac-attenuator -9
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5 | Plac-attenuator -10
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6 | Plac-attenuator -11
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7 | Plac-attenuator -12
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8 | 100bp ladder
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Lane | Sample
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1 | 100bp ladder
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2 | Plac-pT181 antisense
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Colony PCR
Ligation
No Name
state | Vector | Inserter | Ligation High ver.2
|
experiment | 9/6 Plux (SpeI&PstI) | 2 µL | 9/7 RBS-lysis2-DT (EcoRI&XbaI) | µL | 3.5 µL
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experiment | 9/6 Plac (SpeI&PstI) | µL | 9/7 RBS-lysis2-DT (EcoRI&XbaI) | µL | 3.5 µL
|
experiment | 9/6 Pcon-RBS-luxR-DT (EcoRI&XbaI) | 1.9 µL | 9/6 Plux-RBS-GFP-DT(SpeI&PstI) | 2.2 µL | 3.5 µL
|
experiment | 9/6 Pbed/araC (SpeI&PstI) | 1.6 µL | 9/5 RBS-luxI-DT (EcoRI&XbaI) | 16 µL | 3.5 µL
|
experiment | 9/6 Plac (SpeI&PstI) | 1.5 µL | 9/5 pT181 antisense (EcoRI&XbaI) | 2.0 µL | 3.5 µL
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experiment | 9/6 Pcon (SpeI&PstI) | 3.0 µL | 9/5 RBS-luxI-DT (EcoRI&XbaI) | 10 µL | 3.5 µL
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Colony PCR
No name
Sample | base pair
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9/5 Plac-spinarch-DT-13~20 |
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9/5 Plac-pT181 attenuator-13~20 |
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PreDenature | Denature | Annealing | Extension | cycle
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94 °C | 94 °C | 55 °C | 60 °C | --
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5min | 30s | 30s | 42s | 30cycle
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Transformation
Tatsui
Name | Sample | Competent Cells | Total | Plate
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Pcon-RBS-luxR-DT+Plux-RBS-GFP-DT | 2µL | 20µL | 22µL |
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Pbed/araC+RBS-luxI-DT | 2µL | 20µL | 22µL |
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Plac+pT181 antisence | 2µL | 20µL | 22µL |
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Pcon+RBS-luxI-DT | 2µL | 20µL | 22µL |
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Restriction Enzyme Digestion
No Name
| DNA | XbaI | PstI | Buffer | BSA | MilliQ | total
|
9/7 RBS-lysis1-DT(XbaI & PstI) | 10 µL | 1 µL | 1 µL | 3 µL | 3 µL | 12 µL | 30 µL
|
NC | 0.5 µL | 0 µL | 0 µL | 1 µL | 1 µL | 7.5 µL | 10 µL
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| DNA | XbaI | PstI | Buffer | BSA | MilliQ | total
|
9/7 RBS-lysis3-DT (XbaI & PstI) | 7.8 µL | 1 µL | 1 µL | 3 µL | 3 µL | 14.2 µL | 30 µL
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NC | 0.4 µL | 0 µL | 0 µL | 1 µL | 1 µL | 7.6 µL | 10 µL
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| DNA | SpeI | PstI | Buffer | BSA | MilliQ | total
|
9/5 Plac (SpeI & PstI) | 6.0 µL | 1 µL | 1 µL | 3 µL | 3 µL | 16.0 µL | 30 µL
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NC | 0.3 µL | 0 µL | 0 µL | 1 µL | 1 µL | 7.7 µL | 10 µL
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| DNA | EcorI | SpeI | Buffer | BSA | MilliQ | total
|
9/7 pSB4K5 (EcoRI & SpeI) | 8.4 µL | 1 µL | 1 µL | 3 µL | 3 µL | 16.0 µL | 30 µL
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NC | 0.4 µL | 0 µL | 0 µL | 1 µL | 1 µL | 7.7 µL | 10 µL
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| DNA | restricion enzyme | restriction enzyme | Buffer | BSA | MilliQ | total
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Pcon-pT181 attenuator (SpeI & PstI) | 8.6 µL | 1 µL | 1 µL | 3 µL | 3 µL | 13.4 µL | 30 µL
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Pcon-pT181 attenuator (EcorI & SpeI) | 8.6 µL | 1 µL | 1 µL | 3 µL | 3 µL | 13.4 µL | 30 µL
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NC | 0.4 µL | 0 µL | 0 µL | 1 µL | 1 µL | 7.6 µL | 10 µL
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| DNA | XbaI | PstI | Buffer | BSA | MilliQ | total
|
9/5 Spin-DT (XbaI & PstI) | 5.4 µL | 1 µL | 1 µL | 3 µL | 3 µL | 16.6 µL | 30 µL
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NC | 0.3 µL | 0 µL | 0 µL | 1 µL | 1 µL | 7.7 µL | 10 µL
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| DNA | SpeI | PstI | Buffer | BSA | MilliQ | total
|
Ptet-anticodone (SpeI & PstI) | 11 µL | 1 µL | 1 µL | 3 µL | 3 µL | 11 µL | 30 µL
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NC | 0.5 µL | 0 µL | 0 µL | 1 µL | 1 µL | 7.5 µL | 10 µL
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| DNA | XbaI | PstI | Buffer | BSA | MilliQ | total
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9/6 RBS-lacZα-DT (XbaI & PstI) | 11 µL | 1 µL | 1 µL | 3 µL | 3 µL | 11 µL | 30 µL
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NC | 0.5 µL | 0 µL | 0 µL | 1 µL | 1 µL | 7.5 µL | 10 µL
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| DNA | EcoRI | SpeI | Buffer | BSA | MilliQ | total
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pSB1C3 (1) (EcoRI & SpeI) | 6.6 µL | 1 µL | 1 µL | 3 µL | 3 µL | 15.4 µL | 30 µL
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NC | 0.3 µL | 0 µL | 0 µL | 1 µL | 1 µL | 7.7 µL | 10 µL
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| DNA | XbaI | PstI | Buffer | BSA | MilliQ | total
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pSB1C3 (3) (XbaI & PstI) | 6.6 µL | 1 µL | 1 µL | 3 µL | 3 µL | 15.4 µL | 30 µL
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NC | 0.3 µL | 0 µL | 0 µL | 1 µL | 1 µL | 7.7 µL | 10 µL
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