Template:Kyoto/Notebook/Sep 18

From 2013.igem.org

(Difference between revisions)
(→Restriction Enzyme Digestion)
(→Gel Extraction)
Line 205: Line 205:
|1||1kbp ldder||--
|1||1kbp ldder||--
|-
|-
-
|2||Plac②||Spel+Pstl
+
|2||Plac②||SpeI+PstI
|-
|-
-
|3||Plac②||Spel+Pstl
+
|3||Plac②||SpeI+PstI
|-
|-
|4||--||--
|4||--||--
|-
|-
-
|5||RBS-lysis3-DT||Xbal+Pstl
+
|5||RBS-lysis3-DT||XbaI+PstI
|-
|-
-
|6||RBS-lysis3-DT||Xbal+Pstl
+
|6||RBS-lysis3-DT||XbaI+PstI
|-
|-
|7||--||--
|7||--||--
|-
|-
-
|8||Plux-RBS-GFP-DT||EcoR1+Spel
+
|8||Plux-RBS-GFP-DT||EcoRI+SpeI
|-
|-
-
|9||Plux-RBS-GFP-DT||EcoR1+Spel
+
|9||Plux-RBS-GFP-DT||EcoRi+SpeI
|-
|-
|10||--||--
|10||--||--
|-
|-
-
|11||PSB4KS||EcoR1+Spel
+
|11||PSB4KS||EcoRI+SpeI
|-
|-
-
|12||PSB4KS||EcoR1+Spel
+
|12||PSB4KS||EcoRI+SpeI
|}
|}
[[File:igku_9188.jpg]]<br>
[[File:igku_9188.jpg]]<br>
Line 234: Line 234:
|1||1kbp ldder||--
|1||1kbp ldder||--
|-
|-
-
|2||Pcon||Spel+Pstl
+
|2||Pcon||SpeI+PstI
|-
|-
-
|3|| Pcon||Spel+Pstl
+
|3|| Pcon||SpeI+PstI
|-
|-
|4||--||--
|4||--||--
|-
|-
-
|5||Plux||Spel+PStl
+
|5||Plux||SpeI+PStI
|-
|-
-
|6|| Plux||Spel+PStl
+
|6|| Plux||SpeI+PStI
|}
|}
[[File:igku_9184.jpg]]<br>
[[File:igku_9184.jpg]]<br>
Line 249: Line 249:
!Lane||DNA||Enzyme
!Lane||DNA||Enzyme
|-
|-
-
|1||Pcon PT181 antisense spinach-DT||Xbal+Pstl
+
|1||Pcon PT181 antisense spinach-DT||XbaI+PstI
|-
|-
-
|2|| Pcon PT181 antisense Spinach-DT||Xbal+Pstl
+
|2|| Pcon PT181 antisense Spinach-DT||XbaI+PstI
|-
|-
|3||100bp ladder ||--
|3||100bp ladder ||--
|-
|-
-
|4||Pcon PT181 antisense Spinach-DT||Xbal+Pstl
+
|4||Pcon PT181 antisense Spinach-DT||XbaI+PstI
|-
|-
-
|5|| Pcon PT181 antisense Spinach-DT||Xbal+Pstl
+
|5|| Pcon PT181 antisense Spinach-DT||XbaI+PstI
|-
|-
|6|| 1kbp ladder||--
|6|| 1kbp ladder||--
Line 268: Line 268:
|1||100bp ladder ||--
|1||100bp ladder ||--
|-
|-
-
|2||apt12-1R-DT||Xbal+Pstl
+
|2||apt12-1R-DT||XbaI+PstI
|-
|-
-
|3||apt12-1R-DT||Xbal+Pstl
+
|3||apt12-1R-DT||XbaI+PstI
|-
|-
|4||--||--
|4||--||--
|-
|-
-
|5||PT181 attenuator||Xbal+Pstl
+
|5||PT181 attenuator||XbaI+PstI
|-
|-
-
|6||PT181 attenuator||Xbal+Pstl
+
|6||PT181 attenuator||XbaI+PstI
|}
|}
[[File:igku_91810.jpg]]<br>
[[File:igku_91810.jpg]]<br>
Line 285: Line 285:
|1||100bp ladder ||--
|1||100bp ladder ||--
|-
|-
-
|2||Pcoa attenuator||EcoR1+Pstl
+
|2||Pcoa attenuator||EcoRI+PstI
|-
|-
-
|3||Pcoa attenuator||EcoR1+Pstl
+
|3||Pcoa attenuator||EcoRI+PstI
|-
|-
|4||--||--
|4||--||--
|-
|-
-
|5||RBS-lacZ&alpha;-DT||Xbal+Pstl
+
|5||RBS-lacZ&alpha;-DT||XbaI+PstI
|-
|-
-
|6||RBS-lacZ&alpha;-DT ||Xbal+Pstl
+
|6||RBS-lacZ&alpha;-DT ||XbaI+PstI
|}</div>
|}</div>
[[File:igku_91812.jpg]]<br>
[[File:igku_91812.jpg]]<br>
[[File:igku_91813.jpg]]<br>
[[File:igku_91813.jpg]]<br>

Revision as of 00:50, 27 September 2013

Contents

Sep 18

Restriction Enzyme Digestion

No name

9/18Plux SpelPtslBufferBSAMilliQtotal
2cuts3.2µL1µL1µL3µL3µL18.8µL 30µL
NC1.7µL0µL0µL1µL1µL6.3µL 10µL
9/18Pcon-pT181antisense-spinach-DT②XbalPtslBufferBSAMilliQtotal
2cuts7.5µL1µL1µL3µL3µL14.5µL 30µL
NC1.4icro;L0µL0µL1µL1µL6.6µL 10µL
9/16 Plac②SpelPtslBufferBSAMilliQtotal
2cuts13.8µL1µL1µL3µL3µL14.5µL 30µL
NC0.7µL0µL0µL1µL1µL7.3µL 10µL
9/16 Spinach-DTXbalPtslBufferBSAMilliQtotal
2cuts17.6µL1µL1µL3µL3µL4.4µL 30µL
NC0.9µL0µL0µL1µL1µL7.1µL 10µL
9/8 Pcon J23100SpelPtslBufferBSAMilliQtotal
2cuts5.7µL1µL1µL3µL3µL16.3µL 30µL
NC0.3µL0µL0µL1µL1µL7.7µL 10µL
9/16 RBS-lacZα-DTXbalPtslBufferBSAMilliQtotal
2cuts8.4µL1µL1µL3µL3µL13.6µL 30µL
NC0.4µL0µL0µL1µL1µL7.6µL 10µL
9/11 Pcon-pT181antisense-spinach-DT&#9312:XbalPtslBufferBSAMilliQtotal
2cuts7.2µL1µL1µL3µL3µL14.8µL 30µL
NC0.4µL0µL0µL1µL1µL7.6µL 10µL
9/6 apt12-1R-DTXbalPtslBufferBSAMilliQtotal
2cuts7.9µL1µL1µL3µL3µL14.1µL 30µL
NC0.4µL0µL0µL1µL1µL7.6µL 10µL
9/3 pT181antisenseXbalPtslBufferBSAMilliQtotal
2cuts6.3µL1µL1µL3µL3µL15.7µL 30µL
NC0.3µL0µL0µL1µL1µL7.7µL 10µL
9/14 pSB4K5EcoR1SpelBufferBSAMilliQtotal
2cuts7.3µL1µL1µL3µL3µL14.7µL 30µL
NC0.4µL0µL0µL1µL1µL7.6µL 10µL
8/31 Plux-RBS-GFP-DT EcoR1SpelBufferBSAMilliQtotal
2cuts9.2µL1µL1µL3µL3µL12.8µL 30µL
NC0.5µL0µL0µL1µL1µL7.5µL 10µL
9/12 RBS-lysis3-DT XbalPtslBufferBSAMilliQtotal
2cuts5.2µL1µL1µL3µL3µL16.8µL 30µL
NC0.3µL0µL0µL1µL1µL7.6µL 10µL
9/18 Pcon-pT181attenuatorEcoR1SpelBufferBSAMilliQtotal
2cuts14.3µL1µL1µL3µL3µL7.7µL 30µL
NC0.7µL0µL0µL1µL1µL7.3µL 10µL

Electrophoresis

No name

LaneSampleEnzyme1Enzyme2
1Plae② SpelPtsl
2RBS-lysis3-DTXbalPtsl
3Plux-RBS-GFP-DTEcoR1Spel
4PSB4K5EcoR1Spel
5PconSpelPstl
6PluxSpelPstl
71kbp ladder----
8Plae② ----
9RBS-lysis3-DT----
10Plux-RBS-GFP-DT----
11PSB4K5----
12Pcon----
13Plux----

Igku 9183.jpg

Electrophoresis

Okazaki

LaneSampleEnzyme1Enzyme2
1100bp ladder -- --
2Pcon pT181antisense spinach-DT②XbalPstl
3Spinach-DTXbalPstl
4RBS-lacZα-DT XbalPstl
5Pcon pT181antisense-Spinach-DT① XbalPstl
6apt12-1R-DTXbalPstl
7pT181attenuatorXbalPstl
8Pcon attenuatorEcoR1Spel

Igku 9181.jpg

LaneSampleEnzyme1Enzyme2
1100bp ladder -- --
2Pcon pT181antisense spinach-DT②XbalPstl
3Spinach-DTXbalPstl
4RBS-lacZα-DT XbalPstl
5Pcon pT181antisense-Spinach-DT① XbalPstl
6apt12-1R-DTXbalPstl
7pT181attenuatorXbalPstl
8Pcon attenuatorEcoR1Spel
LaneSampleEnzyme1Enzyme2
1100bp ladder -- --
2Pcon pT181antisense spinach-DT②----
3Spinach-DT----
4RBS-lacZα-DT----
5Pcon pT181antisense-Spinach-DT①----
6apt12-1R-DT----
7pT181attenuator----
8Pcon attenuator----

Igku 9182.jpg

Gel Extraction

No name

LaneDNAEnzyme
11kbp ldder--
2Plac②SpeI+PstI
3Plac②SpeI+PstI
4----
5RBS-lysis3-DTXbaI+PstI
6RBS-lysis3-DTXbaI+PstI
7----
8Plux-RBS-GFP-DTEcoRI+SpeI
9Plux-RBS-GFP-DTEcoRi+SpeI
10----
11PSB4KSEcoRI+SpeI
12PSB4KSEcoRI+SpeI

Igku 9188.jpg
Igku 9189.jpg

LaneDNAEnzyme
11kbp ldder--
2PconSpeI+PstI
3 PconSpeI+PstI
4----
5PluxSpeI+PStI
6 PluxSpeI+PStI

Igku 9184.jpg
Igku 9185.jpg

LaneDNAEnzyme
1Pcon PT181 antisense spinach-DTXbaI+PstI
2 Pcon PT181 antisense Spinach-DTXbaI+PstI
3100bp ladder --
4Pcon PT181 antisense Spinach-DTXbaI+PstI
5 Pcon PT181 antisense Spinach-DTXbaI+PstI
6 1kbp ladder--

Igku 9186.jpg
Igku 9187.jpg

LaneDNAEnzyme
1100bp ladder --
2apt12-1R-DTXbaI+PstI
3apt12-1R-DTXbaI+PstI
4----
5PT181 attenuatorXbaI+PstI
6PT181 attenuatorXbaI+PstI

Igku 91810.jpg
Igku 91811.jpg

LaneDNAEnzyme
1100bp ladder --
2Pcoa attenuatorEcoRI+PstI
3Pcoa attenuatorEcoRI+PstI
4----
5RBS-lacZα-DTXbaI+PstI
6RBS-lacZα-DT XbaI+PstI

Igku 91812.jpg
Igku 91813.jpg

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