Template:Kyoto/Notebook/Sep 21

From 2013.igem.org

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===Restriction Enzyme Digestion===
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<div class="experiment">
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<span class="author">Noname</span>
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{| class="wikitable"
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! ||8/17 RBS-DT||EcoRI||SpeI||XbaI||PstI|| Buffer||BSA||MilliQ||total
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|-
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|2cuts(XbaI&PstI)||8.1||0&micro;L||0&micro;L||1&micro;L||1&micro;L||3&micro;L||3&micro;L||13.9&micro;L||30&micro;L
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|-
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|NC||0.4||0&micro;L||0&micro;L||0&micro;L||0&micro;L||1&micro;L||1&micro;L||7.6&micro;L||10&micro;L
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|-
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|}
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{| class="wikitable"
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! ||8/17 RBS-DT||EcoRI||SpeI||XbaI||PstI||Buffer||BSA||MilliQ||total
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|-
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|2cut(EcoRI&XbaI)||16.2||1&micro;L||1&micro;L||0&micro;L||0&micro;L||3&micro;L||3&micro;L||5.8&micro;L||30&micro;L
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|}
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{| class="wikitable"
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! ||9/15 Pcon-pT181attenuator||EcoRI||SpeI||XbaI||PstI|| Buffer||BSA||MilliQ||total
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|-
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|2cuts(EcoRI&PstI)||12||1&micro;L||0&micro;L||0&micro;L||1&micro;L||3&micro;L||3&micro;L||10&micro;L||30&micro;L
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|-
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|NC||0.6||0&micro;L||0&micro;L||0&micro;L||0&micro;L||1&micro;L||1&micro;L||7.4&micro;L||10&micro;L
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|-
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|}
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{| class="wikitable"
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! ||8/20 Pcon=RBS-GFP-DT-1||EcoRI||SpeI||XbaI||PstI||Buffer||BSA||MilliQ||total
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|-
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|2cut(EcoRI&XbaI)||6||1&micro;L||0&micro;L||1&micro;L||0&micro;L||3&micro;L||3&micro;L||0&micro;L||14&micro;L
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|-
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|NC||0.3||0&micro;L||0&micro;L||0&micro;L||0&micro;L||1&micro;L||1&micro;L||7.3&micro;L||10&micro;L
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|}
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incubate 37&deg;C 1hour

Revision as of 14:59, 25 September 2013

Contents

Sep 21

Electrophoresis

じっけんしゃ

LaneSampleEnzyme1Enzyme2
1Pcon-RBS-lacZα-DT----
21kbp ladder----
3pSB1C3+pT181 attenuator----

File:Igku xxxxxx.xxx

Liquid Culture

No name

Samplemedium
9/2 spinach-DT-2--
8/16 Plux-4--
8/27 Plux-RBS-GFP-DT-1--
9/5 Pcon-pT181 attenuator-1--

Master Plate

No name

NumberUse LB plate
18/27 Pλ-luxI-1(+Amp)
29/10 Plac(BBa-R0011)-3(+Amp)
39/5 Pcon-RBS-tetR-DT-1(+Amp)
49/5 Pcon-pT181 antisense-1(+Amp)
59/15 Pcon-pT181 attenuator(+Amp)
69/- Pcon-apt12_1R-DT-1(+Amp)
79/5 Pcon-spinach-DT-3(+Amp)
88/27 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT(+Amp)
NumberUse LB plate
19/4 pT181 antisense(pSB1C3)-1(+CP)
29/11 Fusion1 antisense(pSB1C3)-4(+CP)
39/11 Fusion6 antisense(pSB1C3)-1(+CP)
49/4 pT181 attenuator(pSB1C3)-1(+CP)
59/10 Fusion1 attenuator(pSB1C3)-1(+CP)
69/11 Fusion3n2 attenuator(pSB1C3)-1(+CP)
79/4 aptamer12_1R(pSB1C3)-1(+CP)
89/11 aptamer12_P(pSB1C3)-3(+CP)
99/11 aptamer12_1M(pSB1C3)-1(+CP)

</div>

NumberUse LB plate
18/29 Pbad/araC-RBS-RFP-DT-1(+CP)
29/4 RBS-lysis3-DT-3(+CP)
39/10 Pcon-pT181 attenuator-DT-1(+CP)
49/5 Ptet-pT181 antisense-1(+CP)
59/4aptamer 12_1R-DT-1(+CP)
69/5 Ptet-RBS-lacZα-DT-4(+CP)
78/10 RBS-lacI-DT-2(+CP)
88/29 Plux-RBS-GFP-DT-1(+CP)

</div>

Restriction Enzyme Digestion

Noname

8/17 RBS-DTEcoRISpeIXbaIPstI BufferBSAMilliQtotal
2cuts(XbaI&PstI)8.10µL0µL1µL1µL3µL3µL13.9µL30µL
NC0.40µL0µL0µL0µL1µL1µL7.6µL10µL
8/17 RBS-DTEcoRISpeIXbaIPstIBufferBSAMilliQtotal
2cut(EcoRI&XbaI)16.21µL1µL0µL0µL3µL3µL5.8µL30µL
9/15 Pcon-pT181attenuatorEcoRISpeIXbaIPstI BufferBSAMilliQtotal
2cuts(EcoRI&PstI)121µL0µL0µL1µL3µL3µL10µL30µL
NC0.60µL0µL0µL0µL1µL1µL7.4µL10µL
8/20 Pcon=RBS-GFP-DT-1EcoRISpeIXbaIPstIBufferBSAMilliQtotal
2cut(EcoRI&XbaI)61µL0µL1µL0µL3µL3µL0µL14µL
NC0.30µL0µL0µL0µL1µL1µL7.3µL10µL
incubate 37°C 1hour
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