Template:Kyoto/Notebook/Sep 18

From 2013.igem.org

(Difference between revisions)
(→Gel Extraction)
(→Electrophoresis)
 
(11 intermediate revisions not shown)
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
Line 11: Line 11:
|}
|}
{| class="wikitable"
{| class="wikitable"
-
!||9/18Pcon-pT181antisense-spin-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-spin-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
Line 53: Line 53:
|}
|}
{| 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
Line 60: Line 60:
|}
|}
{| 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
Line 67: Line 67:
|}
|}
{| 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
Line 74: Line 74:
|}
|}
{| 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
Line 81: Line 81:
|}
|}
{| 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
Line 88: Line 88:
|}
|}
{| 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||--||--
Line 129: Line 129:
|13||Plux||--||--
|13||Plux||--||--
|}
|}
-
[[File:igku_xxxxxx.xxx]]<br>
+
[[File:igku_9183.jpg]]<br>
</div>
</div>
Line 141: Line 141:
|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>
{| class="wikitable"
{| class="wikitable"
!Lane||Sample||Enzyme1||Enzyme2
!Lane||Sample||Enzyme1||Enzyme2
Line 160: Line 161:
|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"
Line 193: Line 194:
|8||Pcon attenuator||--||--
|8||Pcon attenuator||--||--
|}
|}
-
[[File:igku_xxxxxx.xxx]]<br>
+
[[File:igku_9182.jpg]]<br>
</div>
</div>
Line 204: Line 205:
|1||1kbp ldder||--
|1||1kbp ldder||--
|-
|-
-
|2||Plac&#9313;||Spel+Pstl
+
|2||Plac&#9313;||SpeI+PstI
|-
|-
-
|3||Plac&#9313;||Spel+Pstl
+
|3||Plac&#9313;||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_xxbeforexx.xxx]]<br>
+
[[File:igku_9188.jpg]]<br>
-
[[File:igku_xxafterxx.xxx]]<br>
+
[[File:igku_9189.jpg]]<br>
{| class="wikitable"
{| class="wikitable"
!Lane||DNA||Enzyme
!Lane||DNA||Enzyme
Line 233: 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_9185.jpg]]<br>
{| class="wikitable"
{| class="wikitable"
!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||--
|}
|}
 +
[[File:igku_9186.jpg]]<br>
 +
[[File:igku_9187.jpg]]<br>
{| class="wikitable"
{| class="wikitable"
!Lane||DNA||Enzyme
!Lane||DNA||Enzyme
Line 263: 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_91811.jpg]]<br>
{| class="wikitable"
{| class="wikitable"
!Lane||DNA||Enzyme
!Lane||DNA||Enzyme
Line 278: 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_91813.jpg]]<br>

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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