Template:Kyoto/Notebook/Sep 18

From 2013.igem.org

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Line 4: Line 4:
<span class="author">No name</span>
<span class="author">No name</span>
{| class="wikitable"
{| class="wikitable"
-
! ||9/18Plux ||Spel||Ptsl||Buffer||BSA||MilliQ||total
+
! ||9/18Plux ||SpeI||PstI||Buffer||BSA||MilliQ||total
|-
|-
|2cuts||3.2&micro;L||1&micro;L||1&micro;L||3&micro;L||3&micro;L||18.8&micro;L|| 30&micro;L
|2cuts||3.2&micro;L||1&micro;L||1&micro;L||3&micro;L||3&micro;L||18.8&micro;L|| 30&micro;L
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|}
|}
{| class="wikitable"
{| class="wikitable"
-
!||9/18Pcon-pT181antisense-spinach-DT&#9313;||Xbal||Ptsl||Buffer||BSA||MilliQ||total
+
!||9/18Pcon-pT181antisense-spinach-DT&#9313;||XbaI||PstI||Buffer||BSA||MilliQ||total
|-
|-
|2cuts||7.5&micro;L||1&micro;L||1&micro;L||3&micro;L||3&micro;L||14.5&micro;L|| 30&micro;L
|2cuts||7.5&micro;L||1&micro;L||1&micro;L||3&micro;L||3&micro;L||14.5&micro;L|| 30&micro;L
Line 18: Line 18:
|}
|}
{| class="wikitable"
{| class="wikitable"
-
! ||9/16 Plac&#9313;||Spel||Ptsl||Buffer||BSA||MilliQ||total
+
! ||9/16 Plac&#9313;||SpeI||PstI||Buffer||BSA||MilliQ||total
|-
|-
|2cuts||13.8&micro;L||1&micro;L||1&micro;L||3&micro;L||3&micro;L||14.5&micro;L|| 30&micro;L
|2cuts||13.8&micro;L||1&micro;L||1&micro;L||3&micro;L||3&micro;L||14.5&micro;L|| 30&micro;L
Line 25: Line 25:
|}
|}
{| class="wikitable"
{| class="wikitable"
-
! ||9/16 Spinach-DT||Xbal||Ptsl||Buffer||BSA||MilliQ||total
+
! ||9/16 Spinach-DT||XbaI||PstI||Buffer||BSA||MilliQ||total
|-
|-
|2cuts||17.6&micro;L||1&micro;L||1&micro;L||3&micro;L||3&micro;L||4.4&micro;L|| 30&micro;L
|2cuts||17.6&micro;L||1&micro;L||1&micro;L||3&micro;L||3&micro;L||4.4&micro;L|| 30&micro;L
Line 32: Line 32:
|}
|}
{| class="wikitable"
{| class="wikitable"
-
! ||9/8 Pcon J23100||Spel||Ptsl||Buffer||BSA||MilliQ||total
+
! ||9/8 Pcon J23100||SpeI||PstI||Buffer||BSA||MilliQ||total
|-
|-
|2cuts||5.7&micro;L||1&micro;L||1&micro;L||3&micro;L||3&micro;L||16.3&micro;L|| 30&micro;L
|2cuts||5.7&micro;L||1&micro;L||1&micro;L||3&micro;L||3&micro;L||16.3&micro;L|| 30&micro;L
Line 39: Line 39:
|}
|}
{| class="wikitable"
{| class="wikitable"
-
! ||9/16 RBS-lacZ&alpha;-DT||Xbal||Ptsl||Buffer||BSA||MilliQ||total
+
! ||9/16 RBS-lacZ&alpha;-DT||XbaI||PstI||Buffer||BSA||MilliQ||total
|-
|-
|2cuts||8.4&micro;L||1&micro;L||1&micro;L||3&micro;L||3&micro;L||13.6&micro;L|| 30&micro;L
|2cuts||8.4&micro;L||1&micro;L||1&micro;L||3&micro;L||3&micro;L||13.6&micro;L|| 30&micro;L
Line 46: Line 46:
|}
|}
{| class="wikitable"
{| class="wikitable"
-
! ||9/11 Pcon-pT181antisense-spinach-DT&#9312:||Xbal||Ptsl||Buffer||BSA||MilliQ||total
+
! ||9/11 Pcon-pT181antisense-spinach-DT&#9312:||XbaI||PstI||Buffer||BSA||MilliQ||total
|-
|-
|2cuts||7.2&micro;L||1&micro;L||1&micro;L||3&micro;L||3&micro;L||14.8&micro;L|| 30&micro;L
|2cuts||7.2&micro;L||1&micro;L||1&micro;L||3&micro;L||3&micro;L||14.8&micro;L|| 30&micro;L
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|}
|}
{| class="wikitable"
{| class="wikitable"
-
! ||9/6 apt12-1R-DT||Xbal||Ptsl||Buffer||BSA||MilliQ||total
+
! ||9/6 apt12-1R-DT||XbaI||PstI||Buffer||BSA||MilliQ||total
|-
|-
|2cuts||7.9&micro;L||1&micro;L||1&micro;L||3&micro;L||3&micro;L||14.1&micro;L|| 30&micro;L
|2cuts||7.9&micro;L||1&micro;L||1&micro;L||3&micro;L||3&micro;L||14.1&micro;L|| 30&micro;L
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|}
|}
{| class="wikitable"
{| class="wikitable"
-
! ||9/3 pT181antisense||Xbal||Ptsl||Buffer||BSA||MilliQ||total
+
! ||9/3 pT181antisense||XbaI||PstI||Buffer||BSA||MilliQ||total
|-
|-
|2cuts||6.3&micro;L||1&micro;L||1&micro;L||3&micro;L||3&micro;L||15.7&micro;L|| 30&micro;L
|2cuts||6.3&micro;L||1&micro;L||1&micro;L||3&micro;L||3&micro;L||15.7&micro;L|| 30&micro;L
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|}
|}
{| class="wikitable"
{| class="wikitable"
-
! ||9/14 pSB4K5||EcoR1||Spel||Buffer||BSA||MilliQ||total
+
! ||9/14 pSB4K5||EcoRI||SpeI||Buffer||BSA||MilliQ||total
|-
|-
|2cuts||7.3&micro;L||1&micro;L||1&micro;L||3&micro;L||3&micro;L||14.7&micro;L|| 30&micro;L
|2cuts||7.3&micro;L||1&micro;L||1&micro;L||3&micro;L||3&micro;L||14.7&micro;L|| 30&micro;L
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|}
|}
{| class="wikitable"
{| class="wikitable"
-
! ||8/31 Plux-RBS-GFP-DT ||EcoR1||Spel||Buffer||BSA||MilliQ||total
+
! ||8/31 Plux-RBS-GFP-DT ||EcoRI||SpeI||Buffer||BSA||MilliQ||total
|-
|-
|2cuts||9.2&micro;L||1&micro;L||1&micro;L||3&micro;L||3&micro;L||12.8&micro;L|| 30&micro;L
|2cuts||9.2&micro;L||1&micro;L||1&micro;L||3&micro;L||3&micro;L||12.8&micro;L|| 30&micro;L
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|}
|}
{| class="wikitable"
{| class="wikitable"
-
! ||9/12 RBS-lysis3-DT ||Xbal||Ptsl||Buffer||BSA||MilliQ||total
+
! ||9/12 RBS-lysis3-DT ||XbaI||PstI||Buffer||BSA||MilliQ||total
|-
|-
|2cuts||5.2&micro;L||1&micro;L||1&micro;L||3&micro;L||3&micro;L||16.8&micro;L|| 30&micro;L
|2cuts||5.2&micro;L||1&micro;L||1&micro;L||3&micro;L||3&micro;L||16.8&micro;L|| 30&micro;L
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|}
|}
{| class="wikitable"
{| class="wikitable"
-
! ||9/18 Pcon-pT181attenuator||EcoR1||Spel||Buffer||BSA||MilliQ||total
+
! ||9/18 Pcon-pT181attenuator||EcoRI||SpeI||Buffer||BSA||MilliQ||total
|-
|-
|2cuts||14.3&micro;L||1&micro;L||1&micro;L||3&micro;L||3&micro;L||7.7&micro;L|| 30&micro;L
|2cuts||14.3&micro;L||1&micro;L||1&micro;L||3&micro;L||3&micro;L||7.7&micro;L|| 30&micro;L
Line 103: Line 103:
!Lane||Sample||Enzyme1||Enzyme2
!Lane||Sample||Enzyme1||Enzyme2
|-
|-
-
|1||Plae&#9313; ||Spel||Ptsl
+
|1||Plae&#9313; ||SpeI||PstI
|-
|-
-
|2||RBS-lysis3-DT||Xbal||Ptsl
+
|2||RBS-lysis3-DT||XbaI||PstI
|-
|-
-
|3||Plux-RBS-GFP-DT||EcoR1||Spel
+
|3||Plux-RBS-GFP-DT||EcoRI||SpeI
|-
|-
-
|4||PSB4K5||EcoR1||Spel
+
|4||PSB4K5||EcoRI||SpeI
|-
|-
-
|5||Pcon||Spel||Pstl
+
|5||Pcon||SpeI||PstI
|-
|-
-
|6||Plux||Spel||Pstl
+
|6||Plux||SpeI||PstI
|-
|-
|7||1kbp ladder||--||--
|7||1kbp ladder||--||--
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|1||100bp ladder ||-- ||--
|1||100bp ladder ||-- ||--
|-
|-
-
|2||Pcon pT181antisense spinach-DT&#9313;||Xbal||Pstl
+
|2||Pcon pT181antisense spinach-DT&#9313;||XbaI||PstI
|-
|-
-
|3||Spinach-DT||Xbal||Pstl
+
|3||Spinach-DT||XbaI||PstI
|-
|-
-
|4||RBS-lacZ&alpha;-DT|| Xbal||Pstl
+
|4||RBS-lacZ&alpha;-DT|| XbaI||PstI
|-
|-
-
|5||Pcon pT181antisense-Spinach-DT&#9312;|| Xbal||Pstl
+
|5||Pcon pT181antisense-Spinach-DT&#9312;|| XbaI||PstI
|-
|-
-
|6||apt12-1R-DT||Xbal||Pstl
+
|6||aptamer12-1R-DT||XbaI||PstI
|-
|-
-
|7||pT181attenuator||Xbal||Pstl
+
|7||pT181attenuator||XbaI||PstI
|-
|-
-
|8||Pcon attenuator||EcoR1||Spel
+
|8||Pcon attenuator||EcoRI||SpeI
|}
|}
[[File:igku_9181.jpg]]<br>
[[File:igku_9181.jpg]]<br>
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|1||100bp ladder ||-- ||--
|1||100bp ladder ||-- ||--
|-
|-
-
|2||Pcon pT181antisense spinach-DT&#9313;||Xbal||Pstl
+
|2||Pcon pT181antisense spinach-DT&#9313;||XbaI||PstI
|-
|-
-
|3||Spinach-DT||Xbal||Pstl
+
|3||Spinach-DT||XbaI||PstI
|-
|-
-
|4||RBS-lacZ&alpha;-DT|| Xbal||Pstl
+
|4||RBS-lacZ&alpha;-DT||XbaI||PstI
|-
|-
-
|5||Pcon pT181antisense-Spinach-DT&#9312;|| Xbal||Pstl
+
|5||Pcon pT181antisense-Spinach-DT&#9312;||XbaI||PstI
|-
|-
-
|6||apt12-1R-DT||Xbal||Pstl
+
|6||apt12-1R-DT||XbaI||PstI
|-
|-
-
|7||pT181attenuator||Xbal||Pstl
+
|7||pT181attenuator||XbaI||PstI
|-
|-
-
|8||Pcon attenuator||EcoR1||Spel
+
|8||Pcon attenuator||EcoRI||SpeI
|}
|}
{| class="wikitable"
{| class="wikitable"

Latest revision as of 07:47, 27 September 2013

Contents

Sep 18

Restriction Enzyme Digestion

No name

9/18Plux SpeIPstIBufferBSAMilliQtotal
2cuts3.2µL1µL1µL3µL3µL18.8µL 30µL
NC1.7µL0µL0µL1µL1µL6.3µL 10µL
9/18Pcon-pT181antisense-spinach-DT②XbaIPstIBufferBSAMilliQtotal
2cuts7.5µL1µL1µL3µL3µL14.5µL 30µL
NC1.4icro;L0µL0µL1µL1µL6.6µL 10µL
9/16 Plac②SpeIPstIBufferBSAMilliQtotal
2cuts13.8µL1µL1µL3µL3µL14.5µL 30µL
NC0.7µL0µL0µL1µL1µL7.3µL 10µL
9/16 Spinach-DTXbaIPstIBufferBSAMilliQtotal
2cuts17.6µL1µL1µL3µL3µL4.4µL 30µL
NC0.9µL0µL0µL1µL1µL7.1µL 10µL
9/8 Pcon J23100SpeIPstIBufferBSAMilliQtotal
2cuts5.7µL1µL1µL3µL3µL16.3µL 30µL
NC0.3µL0µL0µL1µL1µL7.7µL 10µL
9/16 RBS-lacZα-DTXbaIPstIBufferBSAMilliQtotal
2cuts8.4µL1µL1µL3µL3µL13.6µL 30µL
NC0.4µL0µL0µL1µL1µL7.6µL 10µL
9/11 Pcon-pT181antisense-spinach-DT&#9312:XbaIPstIBufferBSAMilliQtotal
2cuts7.2µL1µL1µL3µL3µL14.8µL 30µL
NC0.4µL0µL0µL1µL1µL7.6µL 10µL
9/6 apt12-1R-DTXbaIPstIBufferBSAMilliQtotal
2cuts7.9µL1µL1µL3µL3µL14.1µL 30µL
NC0.4µL0µL0µL1µL1µL7.6µL 10µL
9/3 pT181antisenseXbaIPstIBufferBSAMilliQtotal
2cuts6.3µL1µL1µL3µL3µL15.7µL 30µL
NC0.3µL0µL0µL1µL1µL7.7µL 10µL
9/14 pSB4K5EcoRISpeIBufferBSAMilliQtotal
2cuts7.3µL1µL1µL3µL3µL14.7µL 30µL
NC0.4µL0µL0µL1µL1µL7.6µL 10µL
8/31 Plux-RBS-GFP-DT EcoRISpeIBufferBSAMilliQtotal
2cuts9.2µL1µL1µL3µL3µL12.8µL 30µL
NC0.5µL0µL0µL1µL1µL7.5µL 10µL
9/12 RBS-lysis3-DT XbaIPstIBufferBSAMilliQtotal
2cuts5.2µL1µL1µL3µL3µL16.8µL 30µL
NC0.3µL0µL0µL1µL1µL7.6µL 10µL
9/18 Pcon-pT181attenuatorEcoRISpeIBufferBSAMilliQtotal
2cuts14.3µL1µL1µL3µL3µL7.7µL 30µL
NC0.7µL0µL0µL1µL1µL7.3µL 10µL

Electrophoresis

No name

LaneSampleEnzyme1Enzyme2
1Plae② SpeIPstI
2RBS-lysis3-DTXbaIPstI
3Plux-RBS-GFP-DTEcoRISpeI
4PSB4K5EcoRISpeI
5PconSpeIPstI
6PluxSpeIPstI
71kbp ladder----
8Plae② ----
9RBS-lysis3-DT----
10Plux-RBS-GFP-DT----
11PSB4K5----
12Pcon----
13Plux----

Igku 9183.jpg

Electrophoresis

Okazaki

LaneSampleEnzyme1Enzyme2
1100bp ladder -- --
2Pcon pT181antisense spinach-DT②XbaIPstI
3Spinach-DTXbaIPstI
4RBS-lacZα-DT XbaIPstI
5Pcon pT181antisense-Spinach-DT① XbaIPstI
6aptamer12-1R-DTXbaIPstI
7pT181attenuatorXbaIPstI
8Pcon attenuatorEcoRISpeI

Igku 9181.jpg

LaneSampleEnzyme1Enzyme2
1100bp ladder -- --
2Pcon pT181antisense spinach-DT②XbaIPstI
3Spinach-DTXbaIPstI
4RBS-lacZα-DTXbaIPstI
5Pcon pT181antisense-Spinach-DT①XbaIPstI
6apt12-1R-DTXbaIPstI
7pT181attenuatorXbaIPstI
8Pcon attenuatorEcoRISpeI
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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