Template:Kyoto/Notebook/Sep 5

From 2013.igem.org

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(→Electrophoresis)
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</div>
 
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===Electrophoresis===
 
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<div class="experiment">
 
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<span class="author">No name</span>
 
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{| class="wikitable"
 
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!Lane||Sample||Enzyme1||Enzyme2
 
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|-
 
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|1||100bp ladder||--||--
 
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|-
 
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|2||apt12_1R (pSB1C3)-1||--||--
 
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|-
 
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|3||apt12_1R (pSB1C3)-2||--||--
 
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|-
 
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|4||pT181 antisense(pSB1C3)-1||--||--
 
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|-
 
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|5||pT181 antisense(pSB1C3)-2||--||--
 
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|-
 
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|6||pT181 antisense(pSB1C3)-3||--||--
 
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|-
 
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|7||pT181 antisense(pSB1C3)-4||--||--
 
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|-
 
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|8||100bp ladder||--||--
 
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|}
 
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[[File:igku_xx.xxx]]<br>
 
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</div>
 
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{| class="wikitable"
 
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!Lane||Sample||Enzyme1||Enzyme2
 
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|-
 
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|1||100bp ladder||--||--
 
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|-
 
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|2||apt12_1R-DT-1||--||--
 
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|-
 
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|3||apt12_1R-DT-2||--||--
 
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|-
 
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|4||apt12_1R-DT-3||--||--
 
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|-
 
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|5||apt12_1R-DT-4||--||--
 
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|-
 
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|6||pT181 attenuator(pSB1C3)-1||--||--
 
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|-
 
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|7||pT181 attenuator(pSB1C3)-2||--||--
 
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|-
 
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|8||RBS-lysis1-DT-1||--||--
 
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|-
 
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|9||RBS-lysis1-DT-2||--||--
 
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|-
 
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|10||100bp ladder||--||--
 
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|}
 
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[[File:igku_xx.xxx]]<br>
 
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</div>
 
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{| class="wikitable"
 
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!Lane||Sample||Enzyme1||Enzyme2
 
-
|-
 
-
|1||100bp ladder||--||--
 
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|-
 
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|2||RBS-lysis2-DT-1||--||--
 
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|-
 
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|3||RBS-lysis2-DT-2||--||--
 
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|-
 
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|4||RBS-lysis2-DT-3||--||--
 
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|-
 
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|5||RBS-lysis2-DT-4||--||--
 
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|-
 
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|6||RBS-lysis3-DT-1||--||--
 
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|-
 
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|7||RBS-lysis3-DT-2||--||--
 
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|-
 
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|8||100bp ladder||--||--
 
-
|}
 
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[[File:igku_xx.xxx]]<br>
 
</div>
</div>

Revision as of 09:39, 25 September 2013

Contents

Sep 5

Electrophoresis

No name

LaneSampleEnzyme1Enzyme2
1100bp ladder----
2J23100-3----
3J23100-4----
4J23100-5----
5100bp ladder----

File:Igku xx.xxx

Miniprep

Nakamoto

DNAconcentration[µg/mL]260/280260/230
8/3 J23100-3349.11.691.851
8/10 Plac-2331.31.841.42

Ligation

Kojima

stateVectorInserterLigation High ver.2
experimentPconst13.2spinach-DT2.13.5
experimentPlac15.3spinach-DT2.13.5
experimentPconst13.2pT181 antisense (XbaI & PstI)6.63.5
experimentPlac15.3pT181 antisense (XbaI & PstI)6.63.5
experimentPtet30.1pT181 antisense (XbaI & PstI)6.63.5
experimentPconst13.2pT181 attenuator11.73.5
experimentPlac15.3pT181 attenuator11.73.5
experimentPtet30.1RBS-lacZα-DT3.03.5

incubate 16 °C 1 hour

Electrophoresis

Gel Extraction

No name

LaneDNAEnzyme
11kbp ladder
2RBS-luxI-DT
3RBS-luxI-DT
41kbp ladder
5Pbad/araC
6Pbad/araC

File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx

Nameconcentration[µg/mL]260/280260/230
RBS-luxI-DT23.20.837.89
Pbad/araC51.81.200.11

Colony PCR

No name

Samplebase pair
9/4 apt12_1R(pSB1C3)-1384
9/4 apt12_1R(pSB1C3)-2384
9/4 pT181 antisense(pSB1C3)-1415
9/4 pT181 antisense(pSB1C3)-2415
9/4 pT181 antisense(pSB1C3)-3415
9/4 pT181 antisense(pSB1C3)-4415
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s30s30cycles
Samplebase pair
9/4 apt12_1R-DT-1521
9/4 apt12_1R-DT-2521
9/4 apt12_1R-DT-3521
9/4 apt12_1R-DT-4521
9/4 pT181 attenuator(pSB1C3)601
9/4 pT181 attenuator(pSB1C3)601
9/4 RBS-lysis1-DT613
9/4 RBS-lysis1-DT613
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s36s30cycles
Samplebase pair
9/4 RBS-lysis2-DT-1985
9/4 RBS-lysis2-DT-2985
9/4 RBS-lysis2-DT-3985
9/4 RBS-lysis2-DT-4985
9/4 RBS-lysis3-DT-11210
9/4 RBS-lysis3-DT-21210
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s72s30cycles

Liquid Culture

Nakamoto

Samplemedium
8/9 J23100-5Plusgrow medium(+Amp)

37°C

Transformation

No name

NameSample(µL)Competent Cells(µL)Total(µL)Plate
9/5 Pconst+spinach-DT22022--
9/5 Plac+spinach-DT22022--
9/5 Pconst+pT181 antisense22022--
9/5 Plac+pT181 antisense22022--
9/5 Ptet+pT181 antisense22022--
9/5 Plux+pT181 attenuater22022--
9/5 Ptet+pT181 attenuater22022--
9/5 Ptet+pT181 attenuater22022--
9/5 Pconst+RBS-tetR+DT22022--
9/16(2012) T7-His-FT22022--
9/16(2012) pBr32222022--

Restriction Enzyme Digestion

No name

9/5 Pbad/araCESXPBSABufferMilliQtotal
2 cuts4µL 0µL 1µL 0µL 1µL 3µL 3µL 18µL 30µL
9/5 RBS-luxI-DTESXPBSABufferMilliQtotal
2 cuts4µL 0µL 0µL 1µL 1µL 3µL 3µL18µL 30µL
8/28 PluxESXPBSABufferMilliQtotal
2 cuts 12µL 0µL 1µL 0µL 1µL 3µL 3µL10µL30µL
NC 0.6µL0µL0µL 0µL 0µL1µL1µL7.4µL10µL
9/5 Pconst(J23100)ESXPBSABufferMilliQtotal
2 cuts5.7µL0µL1µL0µL1µL3µL3µL16.3µL30µL
NC 0.3µL0µL0µL 0µL 0µL1µL1µL7.7µL10µL
9/5 PlacESXPBSABufferMilliQtotal
2 cuts6.0µL0µL1µL0µL1µL3µL3µL16µL30µL
NC 0.3µL0µL0µL 0µL 0µL1µL1µL7.7µL10µL
8/31 Plux-RBS-GFP-DT-1ESXPBSABufferMilliQtotal
2 cuts9.4µL1µL1µL0µL0µL3µL3µL12.6µL30µL
NC 0.5µL0µL0µL 0µL 0µL1µL1µL7.5µL10µL
8/20 Pconst-RBS-luxR-DTESXPBSABufferMilliQtotal
2 cuts4.5µL1µL0µL1µL0µL3µL3µL13.5µL30µL
NC 0.2µL0µL0µL 0µL 0µL1µL1µL7.8µL10µL
DNAconcentration[µg/mL]260/280260/230
9/5 RBS-luxI-DT7.61.56-1.42
9/5 Pbad/araC7.51.543.84

Electrophoresis

No name

LaneSampleEnzyme1Enzyme2
11kbp ladder----
2PconstSpeIPstI
3Pconst----
4PlacSpeIPstI
5Plac----
6PluxSpeIPstI
7Plux----
8Pconst-RBS-luxR-DTEcoRIXbaI
9Pconst-RBS-luxR-DT----
10Plux-RBS-GFP-DTEcoRISpeI
11Plux-RBS-GFP-DT----
121kbp ladder----

File:Igku xx.xxx

5*M9(EDTA)

Hirano

volume10ml
Na2HPO4 60mg
KH2PO430mg
NaCl5mg
NH4Cl10mg
Fe(3)-EDTA1263.27mg　

121°C 20minute autoclave

Gel Extraction

No name

LaneDNAEnzyme
11kbp ladder
2PconstSpeI & PstI
3PconstSpeI & PstI

File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx

Nameconcentration[µg/mL]260/280260/230
Pconst5.31.980.06
LaneDNAEnzyme
1100bp ladder
2PlacSpeI & PstI
3PlacSpeI & PstI
5PluxSpeI & PstI
6PluxSpeI & PstI

File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx

Nameconcentration[µg/mL]260/280260/230
Plac (SpeI & PstI)5.21.820.36
Plux(SpeI & PstI)8.01.870.25
LaneDNAEnzyme
11kbp ladder
2Pconst-RBS-luxR-DTEcoRI & XbaI
3Pconst-RBS-luxR-DTEcoRI & XbaI
5Plux-RBS-GFP-DTEcoRI & SpeI
6Plux-RBS-GFP-DTEcoRI & SpeI

File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx

Nameconcentration[µg/mL]260/280260/230
Pconst-RBS-luxR-DT(EcoRI & XbaI)30.61.841.16
Plux-RBS-GFP-DT (EcoRI & SpeI)21.81.870.98

Miniprep

No name

DNAconcentration[µg/mL]260/280260/230
8/9 J23100-5243.01.901.81

Colony PCR

No name

Samplebase pair
9/4 RBS-lysis3-DT-31210
9/4 RBS-lysis3-DT-41210
9/4 RBS-lysis3-DT-51210
9/4 RBS-lysis1-DT-3613
9/4 Ptrc KaiC-1--
9/4 pSB4K51370
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5m30s30s1m 25s30cycles

Electrophoresis

No name

LaneSampleEnzyme1Enzyme2
1100bp ladder----
2RBS-lysis3-DT-3----
3RBS-lysis3-DT-4----
4RBS-lysis3-DT-5----
5RBS-lysis1-DT-3----
6Ptrc KaiC-1----
7pSB4K5----
8100bp ladder----

File:Igku xxxxxx.xxx

Liquid Culture

Hirano

Samplemedium
9/4 apt12_1R-DT-1Plusgrow medium
9/4 pT181 attenuator(pSB1C3)-1Plusgrow medium
9/4 RBS-lysis2-DT-1Plusgrow medium
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