Template:Kyoto/Notebook/Sep 21

From 2013.igem.org

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(→Gel Extraction)
 
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===Electrophoresis===
===Electrophoresis===
<div class="experiment">
<div class="experiment">
-
<span class="author">じっけんしゃ</span>
+
<span class="author">no name</span>
{| class="wikitable"
{| class="wikitable"
!Lane||Sample||Enzyme1||Enzyme2
!Lane||Sample||Enzyme1||Enzyme2
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|3||pSB1C3+pT181 attenuator||--||--
|3||pSB1C3+pT181 attenuator||--||--
|}
|}
-
[[File:igku_xxxxxx.xxx]]<br>
+
[[File:igku_9211.jpg]]<br>
</div>
</div>
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|}  
|}  
</div>
</div>
 +
 +
===Restriction Enzyme Digestion===
 +
<div class="experiment">
 +
<span class="author">Noname</span>
 +
{| class="wikitable"
 +
! ||8/17 RBS-DT||EcoRI||SpeI||XbaI||PstI|| Buffer||BSA||MilliQ||total
 +
|-
 +
|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
 +
|-
 +
|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
 +
|-
 +
|}
 +
 +
{| class="wikitable"
 +
! ||8/17 RBS-DT||EcoRI||SpeI||XbaI||PstI||Buffer||BSA||MilliQ||total
 +
|-
 +
|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
 +
|}
 +
 +
{| class="wikitable"
 +
! ||9/15 Pcon-pT181attenuator||EcoRI||SpeI||XbaI||PstI|| Buffer||BSA||MilliQ||total
 +
|-
 +
|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
 +
|-
 +
|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
 +
|-
 +
|}
 +
 +
{| class="wikitable"
 +
! ||8/20 Pcon=RBS-GFP-DT-1||EcoRI||SpeI||XbaI||PstI||Buffer||BSA||MilliQ||total
 +
|-
 +
|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
 +
|-
 +
|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
 +
|}
 +
incubate 37&deg;C 1hour</div>
 +
 +
===Electrophoresis===
 +
<div class="experiment">
 +
<span class="author">No name</span>
 +
{| class="wikitable"
 +
!Lane||Sample||Enzyme1||Enzyme2
 +
|-
 +
|1||RBS-GFP-DT||XbaI||PstI
 +
|-
 +
|2||RBS-GFP-DT||--||--
 +
|-
 +
|3||RBS-GFP-DT||EcoRI||XbaI
 +
|-
 +
|4||1kbp ladder||--||--
 +
|-
 +
|5||Pcon-pT181attenuator||SpeI||PstI
 +
|-
 +
|6||Pcon-pT181attenuator||--||--
 +
|-
 +
|7||Pcon-RBs-GFP-DT||EcoRI||XbaI
 +
|-
 +
|8||Pcon-RBs-GFP-DT||--||--
 +
|}
 +
[[File:Igku_9212.jpg]]<br>
 +
</div>
 +
 +
===PCR===
 +
<div class="experiment">
 +
<span class="author">Nakamoto</span>
 +
{| class="wikitable"
 +
!Sample||base pair
 +
|-
 +
|Pcon-apt12_1R-DT(E-1A)||50
 +
|-
 +
|Pcon-spinach-DT(E-1A)||402
 +
|-
 +
|Pcon-pT181 attenuator(E-1A)||400
 +
|-
 +
|DT(1-3)||65
 +
|-
 +
|tetR aptamer 12_1R-DT(1-3)||283
 +
|-
 +
|Pcon-pT181 attenuator(1-SA)||303
 +
|-
 +
|spinach-DT(2-1)||338
 +
|-
 +
|Pcon-pT181 attenuator(E-0)||403
 +
|-
 +
|Pcon-pT181 attenuator(2-SA)||403
 +
|-
 +
|spinach-DT(3-2)||395
 +
|-
 +
|Ptet-PT181 anisense ||400
 +
|}
 +
{| class="wikitable"
 +
!PreDenature||Denature||Annealing||Extension||cycle
 +
|-
 +
|94&deg;C||98&deg;C||57&deg;C||68&deg;C||--
 +
|-
 +
|2min||10s||30s||35s||30cycles
 +
|}
 +
 +
===Gel Extraction===
 +
<div class="experiment">
 +
<span class="author">No name</span>
 +
{| class="wikitable"
 +
!Lane||DNA||Enzyme
 +
|-
 +
|1||1kb ladder||--
 +
|-
 +
|3||RBS-GFP-DT||XbaI&PstI
 +
|-
 +
|4||RBS-GFP-DT||XbaI&PstI
 +
|-
 +
|5||RBS-GFP-DT||XbaI&PstI
 +
|}
 +
[[File:igku_9213.jpg]]<br>
 +
[[File:igku_9214.jpg]]<br>
 +
{| class="wikitable"
 +
!Name||concentration[&micro;g/mL]||260/280||260/230
 +
|-
 +
|RBS-GFP-DT(XbaI&PstI)||14.8||1.74||0.47
 +
|-
 +
|RBS-GFP-DT(EcoRI&SpeI)||88.4||1.85||1.49
 +
|-
 +
|Pcon-RBS-GFP-DT(EcoRI&XbaI)||47.5||1.82||1.12
 +
|}

Latest revision as of 01:33, 27 September 2013

Contents

Sep 21

Electrophoresis

no name

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

Igku 9211.jpg

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

Electrophoresis

No name

LaneSampleEnzyme1Enzyme2
1RBS-GFP-DTXbaIPstI
2RBS-GFP-DT----
3RBS-GFP-DTEcoRIXbaI
41kbp ladder----
5Pcon-pT181attenuatorSpeIPstI
6Pcon-pT181attenuator----
7Pcon-RBs-GFP-DTEcoRIXbaI
8Pcon-RBs-GFP-DT----

Igku 9212.jpg

PCR

Nakamoto

Samplebase pair
Pcon-apt12_1R-DT(E-1A)50
Pcon-spinach-DT(E-1A)402
Pcon-pT181 attenuator(E-1A)400
DT(1-3)65
tetR aptamer 12_1R-DT(1-3)283
Pcon-pT181 attenuator(1-SA)303
spinach-DT(2-1)338
Pcon-pT181 attenuator(E-0)403
Pcon-pT181 attenuator(2-SA)403
spinach-DT(3-2)395
Ptet-PT181 anisense 400
PreDenatureDenatureAnnealingExtensioncycle
94°C98°C57°C68°C--
2min10s30s35s30cycles

Gel Extraction

No name

LaneDNAEnzyme
11kb ladder--
3RBS-GFP-DTXbaI&PstI
4RBS-GFP-DTXbaI&PstI
5RBS-GFP-DTXbaI&PstI

Igku 9213.jpg
Igku 9214.jpg

Nameconcentration[µg/mL]260/280260/230
RBS-GFP-DT(XbaI&PstI)14.81.740.47
RBS-GFP-DT(EcoRI&SpeI)88.41.851.49
Pcon-RBS-GFP-DT(EcoRI&XbaI)47.51.821.12
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