Template:Kyoto/Notebook/Aug 27

From 2013.igem.org

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[[File:igku_xxxxxx.xxx]]<br>
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===Gel Extraction===
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<!-- こっから -->
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<div class="experiment">
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<span class="author">Inoue</span>
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{| class="wikitable"
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!Lane||DNA||Enzyme
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|-
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|1||100bpladder||--
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|-
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|2||pT181 attenuator-1||EcoRI&SpeI
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|-
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|3||pT181 attenuator-1||EcoRI&SpeI
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|-
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|4||DT-1||EcoRI&XbaI
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|-
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|5||DT-1||EcoRI&XbaI
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|-
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|6||Pcon-RBS-luxR-DT-2||EcoRI&SpeI
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|-
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|7||Pcon-RBS-luxR-DT-2||EcoRI&SpeI
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|-
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|8||100bpladder||--
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|}
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[[File:igku_xxbeforexx.xxx]]<br>
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[[File:igku_xxafterxx.xxx]]<br>
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{| class="wikitable"
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!Name||quantity||concentration[&micro;g/mL]||260/280||260/230
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|-
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|pT181 attenuator-1(EcoRI&SpeI)||4.2||1.62||0.29||
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|-
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|DT-1(EcoRI&XbaI)||17||1.88||0.88||
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|}
</div>
</div>
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Revision as of 05:19, 19 September 2013

Contents

Aug 27

Restriction Enzyme Digestion

No name

8/21 pT181 attenuator-1(330µg/mL)EcoRISpeI10x BufferB100x BSAMilliQtotal
3.0µL0.5µL0.5µL3.0µL0.3µL22.7µL30µL
0.6µL0µL0µL1µL0.1µL8.3µL10µL
8/17 DT-1(188µg/mL)EcoRIXbaI10x BufferB100x BSAMilliQtotal
3.0µL0.5µL0.5µL3.0µL0.3µL22.7µL30µL
1.0µl0µL0µL1µL0.1µL7.9µL10µL
8/20 Pcon-RBS-luxR-DT-2(344µg/mL)EcoRISpeI10x BufferB100x BSAMilliQtotal
2.9µL0.5µL0.5µL3.0µL0.3µL22.8µL30µL
0.6µl0µL0µL1µL0.1µL8.3µL10µL

Electrophoresis

No name

LaneSampleEnzyme1Enzyme2
1100bp ladder----
28/21 pT181 attenuatorEcoISpeI
38/21 pT181 atteniator----
48/17 DT-1EcoRIXbaI
58/17 DT-1----
68/20 Pcon-RBS-luxR-DT-2EcoISpeI
78/20 Pcon-RBS-luxR-DT-2----
8100bp ladder----

Colony PCR

Kojima and Yoshida

Samplebase pair
8/26 Pλ-luxI(1)
8/26 Pλ-luxI(2)
NC
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s3min30cycles

Liquid Culture

No name

Samplemedium
8/26 Pλ-RBS-luxI-DT-1Plusgrow medium (+Amp)
8/26 Pλ-RBS-luxI-DT-2Plusgrow medium (+Amp)　

Electrophoresis

Hirano

LaneSampleEnzyme1Enzyme2
1100bp ladder----
2pT181 attenuatorEcoR1SpeI
3pT181 attenuator----
4DT-(1)EcoRIXbaI
5DT-(1)----
6Pcon-RBS-luxR-DT-(2)EcoRISpeI
7Pcon-RBS-luxR-DT-(2)----
8100bp ladder----

File:Igku xxxxxx.xxx

Gel Extraction

Inoue

LaneDNAEnzyme
1100bpladder--
2pT181 attenuator-1EcoRI&SpeI
3pT181 attenuator-1EcoRI&SpeI
4DT-1EcoRI&XbaI
5DT-1EcoRI&XbaI
6Pcon-RBS-luxR-DT-2EcoRI&SpeI
7Pcon-RBS-luxR-DT-2EcoRI&SpeI
8100bpladder--

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

Namequantityconcentration[µg/mL]260/280260/230
pT181 attenuator-1(EcoRI&SpeI)4.21.620.29
DT-1(EcoRI&XbaI)171.880.88
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