Template:Kyoto/Notebook/Sep 20

From 2013.igem.org

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(→Electrophoresis)
 
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!Name||Sample||Competent Cells||Plate
!Name||Sample||Competent Cells||Plate
|-
|-
-
|9/17 IC3-Pcon-pT181attenuator-DT||2µL||20µL||CP
+
|9/17 pSB1C3-Pcon-pT181 attenuator-DT||2 µL||20 µL||CP
|-
|-
-
|9/17 IC3-Pcon-aptamer12-1R-DT||2µL||20µL||CP
+
|9/17 pSB1C3-Pcon-aptamer 12_1R-DT||2 µL||20 µL||CP
|-
|-
-
|9/17 IC3-Pcon-antisense-spinach-DT||2µL||20µL||CP
+
|9/17 pSB1C3-Pcon-antisense-spinach-DT||2 µL||20 µL||CP
|-
|-
-
|9/17 IC3-Pcon-RBS-tetR-DTx2||2µL||20µL||CP
+
|9/17 pSB1C3-Pcon-RBS-tetR-DT x2||2 µL||20 µL||CP
|-
|-
-
|9/14 IC3-PkaiBC||2µL||20µL||CP
+
|9/14 pSB1C3-PkaiBC||2 µL||20 µL||CP
|-
|-
-
|9/14 IC3-RpaB||2µL||20µL||CP
+
|9/14 pSB1C3-RpaB||2 µL||20 µL||CP
|-
|-
-
|9/16 IC3-pT181attenuator(EcoRI&SpeI) ||2µL||20µL||CP
+
|9/16 pSB1C3-pT181 attenuator(EcoRI&SpeI) ||2 µL||20 µL||CP
|-
|-
-
|9/17 IC3-pT181attenuator(XbaI&PstI) ||2µL||20µL||CP
+
|9/17 pSB1C3-pT181 attenuator(XbaI&PstI) ||2 µL||20 µL||CP
|-
|-
-
|9/17IC3-Pcon-spinach-DT||2µL||20µL||CP
+
|9/17 pSB1C3-Pcon-spinach-DT||2 µL||20 µL||CP
|-
|-
-
|9/14 RBS-GFP-DT-PkaiBC||2µL||20µL||CP
+
|9/14 RBS-GFP-DT-PkaiBC||2 µL||20 µL||CP
|-
|-
-
|Plux-RBS-lysis3-DT||2µL||20µL||CP
+
|Plux-RBS-lysis3-DT||2 µL||20 µL||CP
|-
|-
-
|9/20 Ptet-pT181antisense-spinach-DT||2µL||20µL||CP
+
|9/20 Ptet-pT181 antisense-spinach-DT||2 µL||20 µL||CP
|-
|-
-
|9/20 Pcon-spinach-DT(4K5) ||2µL||20µL||K
+
|9/20 Pcon-spinach-DT(pSB4K5) ||2 µL||20 µL||Kan
|-
|-
-
|9/20 Pcon-pT181antisense-spinach-DT(4K5) ||2µL||20µL||K
+
|9/20 Pcon-pT181 antisense-spinach-DT(pSB4K5) ||2 µL||20 µL||Kan
|-
|-
-
|9/20 KaiABC||2µL||20µL||K
+
|9/20 KaiABC||2 µL||20 µL||Kan
|-
|-
-
|9/20 KaiABC||2µL||20µL||K
+
|9/20 KaiABC||2 µL||20 µL||Kan
|-
|-
-
|9/20 Plac-RBS-lysis3-DT||2µL||20µL||Amp
+
|9/20 Plac-RBS-lysis3-DT||2 µL||20 µL||Amp
|-
|-
-
|9/20 Plac-aptamer12-1R||2µL||20µL||Amp
+
|9/20 Plac-aptamer 12_1R||2 µL||20 µL||Amp
|-
|-
-
|9/20 Plux-RBS-GFP-DT-Pcon-RBS-luxR-DT||2µL||20µL||Amp
+
|9/20 Plux-RBS-GFP-DT-Pcon-RBS-luxR-DT||2 µL||20 µL||Amp
|-
|-
-
|Plac-aptamer12-1R-DT||2µL||20µL||Amp
+
|Plac-aptamer12_1R-DT||2 µL||20 µL||Amp
|-
|-
-
|9/20 Plac-aptamer12-1R-DT||2µL||20µL||Amp
+
|9/20 Plac-aptamer12_1R-DT||2 µL||20 µL||Amp
|-
|-
-
|9/20Pcon-pT181attenuator-aptamer-DT||2µL||20µL||Amp
+
|9/20Pcon-pT181attenuator-aptamer-DT||2 µL||20 µL||Amp
|-
|-
-
|9/17 Plac-pT181-attenuatorx2||2µL||20µL||Amp
+
|9/17 Plac-pT181-attenuatorx2||2 µL||20 µL||Amp
|-
|-
-
|9/15 Pcon-RBS-lacZα-DT||2µL||20µL||Amp
+
|9/15 Pcon-RBS-lacZα-DT||2 µL||20 µL||Amp
|-
|-
-
|9/14 Pcon-pT181attenuator-RBS-lacZα-DT||2µL||20µL||Amp
+
|9/14 Pcon-pT181attenuator-RBS-lacZα-DT||2 µL||20 µL||Amp
|-
|-
-
|PUC19||2µL||20µL||Amp
+
|PUC19||2 µL||20 µL||Amp
-
|-
+
-
 
+
|}
|}
</div>
</div>
===PCR===
===PCR===
-
<!-- ここから -->
 
<div class="experiment">
<div class="experiment">
<span class="author">Tatsui</span>
<span class="author">Tatsui</span>
Line 88: Line 85:
!Sample||base pair
!Sample||base pair
|-
|-
-
|pT181attenuator(IC3)(XbaI&PstI)||--
+
|pT181 attenuator(pSB1C3)||--
|-
|-
|Pcon-RBS-lacZ&alpha;-DT||--
|Pcon-RBS-lacZ&alpha;-DT||--
Line 95: Line 92:
!PreDenature||Denature||Annealing||Extension||cycle
!PreDenature||Denature||Annealing||Extension||cycle
|-
|-
-
|94&deg;C||94&deg;C||55&deg;C||68&deg;C||--
+
|94 &deg;C||94 &deg;C||55 &deg;C||68 &deg;C||--
|-
|-
-
|5min||30s||30s||3min||30cycles
+
|5 min||30 s||30 s||3 min||30 cycles
|}
|}
</div>
</div>
Line 109: Line 106:
|1||Pcon-RBS-lacZ&alpha;-DT||--
|1||Pcon-RBS-lacZ&alpha;-DT||--
|-
|-
-
|2|| 1kb ladder||--  
+
|2||1kb ladder||--  
|-
|-
-
|3||IC3-pT181attenuator ||--
+
|3||pSB1C3-pT181attenuator ||--
|}
|}
</div>
</div>

Latest revision as of 11:39, 24 September 2013

Contents

Sep 20

Ligation

Okazaki, Kanaoka, Tatsui and Hirano

stateVectorInserterLigation High ver.2
experiment9/18 Plac (SpeI & PstI)3.0 µL9/18 RBS-lysis3-DT (XbaI & PstI)12.4 µL3.5 µL
experiment9/12 Ptet-pT181 antisense1.0 µL9/18 spinach-DT11.9 µL3.5 µL
experiment9/13 Pcon-RBS-luxR-DT2.1 µL9/10 Plux-RBS-GFP-DT5.6 µL3.5 µL
experiment9/13 Pcon-pT181 attenuator (SpeI & PstI)2.0 µL9/18 aptamer 12-1R-DT (XbaI & PstI)6.3 µL3.5 µL
experiment9/18 Plac ②(SpeI & PstI)3.0 µL9/18 aptamer 12-1R-DT(XbaI & PstI)4.5µLx3.5 µL

Transformation

Tatsui

NameSampleCompetent CellsPlate
9/17 pSB1C3-Pcon-pT181 attenuator-DT2 µL20 µLCP
9/17 pSB1C3-Pcon-aptamer 12_1R-DT2 µL20 µLCP
9/17 pSB1C3-Pcon-antisense-spinach-DT2 µL20 µLCP
9/17 pSB1C3-Pcon-RBS-tetR-DT x22 µL20 µLCP
9/14 pSB1C3-PkaiBC2 µL20 µLCP
9/14 pSB1C3-RpaB2 µL20 µLCP
9/16 pSB1C3-pT181 attenuator(EcoRI&SpeI) 2 µL20 µLCP
9/17 pSB1C3-pT181 attenuator(XbaI&PstI) 2 µL20 µLCP
9/17 pSB1C3-Pcon-spinach-DT2 µL20 µLCP
9/14 RBS-GFP-DT-PkaiBC2 µL20 µLCP
Plux-RBS-lysis3-DT2 µL20 µLCP
9/20 Ptet-pT181 antisense-spinach-DT2 µL20 µLCP
9/20 Pcon-spinach-DT(pSB4K5) 2 µL20 µLKan
9/20 Pcon-pT181 antisense-spinach-DT(pSB4K5) 2 µL20 µLKan
9/20 KaiABC2 µL20 µLKan
9/20 KaiABC2 µL20 µLKan
9/20 Plac-RBS-lysis3-DT2 µL20 µLAmp
9/20 Plac-aptamer 12_1R2 µL20 µLAmp
9/20 Plux-RBS-GFP-DT-Pcon-RBS-luxR-DT2 µL20 µLAmp
Plac-aptamer12_1R-DT2 µL20 µLAmp
9/20 Plac-aptamer12_1R-DT2 µL20 µLAmp
9/20Pcon-pT181attenuator-aptamer-DT2 µL20 µLAmp
9/17 Plac-pT181-attenuatorx22 µL20 µLAmp
9/15 Pcon-RBS-lacZα-DT2 µL20 µLAmp
9/14 Pcon-pT181attenuator-RBS-lacZα-DT2 µL20 µLAmp
PUC192 µL20 µLAmp

PCR

Tatsui

Samplebase pair
pT181 attenuator(pSB1C3)--
Pcon-RBS-lacZα-DT--
PreDenatureDenatureAnnealingExtensioncycle
94 °C94 °C55 °C68 °C--
5 min30 s30 s3 min30 cycles

Electrophoresis

No name

LaneSampleEnzyme
1Pcon-RBS-lacZα-DT--
21kb ladder--
3pSB1C3-pT181attenuator --
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