Template:Kyoto/Notebook/Sep 26

From 2013.igem.org

(Difference between revisions)
(→Liquid Culture)
(→Electrophoresis)
Line 6: Line 6:
!Lane||Sample||Enzyme1||Enzyme2
!Lane||Sample||Enzyme1||Enzyme2
|-
|-
-
|1||GGA1-1|| || ||  
+
|1||GGA1-1|| ||  
|-
|-
-
|2||GGA1-2|| || ||  
+
|2||GGA1-2|| ||  
|-
|-
-
|3||GGA1-3|| || ||  
+
|3||GGA1-3|| ||  
|-
|-
-
|4||GGA1-4|| || ||  
+
|4||GGA1-4|| ||  
|-
|-
-
|5||GGA1-5|| || ||  
+
|5||GGA1-5|| ||  
|-
|-
-
|6||GGA1-6|| || ||  
+
|6||GGA1-6|| ||  
|-
|-
-
|7||GGA1-7|| || ||  
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|7||GGA1-7|| ||  
|-
|-
-
|8||GGA1-8|| || ||  
+
|8||GGA1-8|| ||  
|-
|-
-
|9||1kbp ladder|| || ||  
+
|9||1kbp ladder|| ||  
|-
|-
-
|10||GGA2-1|| || ||  
+
|10||GGA2-1|| ||  
|-
|-
-
|11||GGA2-2|| || ||  
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|11||GGA2-2|| ||  
|-
|-
-
|12||GGA2-3|| || ||  
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|12||GGA2-3|| ||  
|-
|-
-
|13||GGA2-4|| || ||  
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|13||GGA2-4|| ||  
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|-
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|14||GGA2-5|| || ||  
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|14||GGA2-5|| ||  
|-
|-
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|15||GGA2-6|| || ||  
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|15||GGA2-6|| ||  
|-
|-
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|16||GGA2-7|| || ||  
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|16||GGA2-7|| ||  
|-
|-
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|17||GGA2-8|| || ||  
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|17||GGA2-8|| ||  
|}
|}
[[File:xxxxx.png]]<br>
[[File:xxxxx.png]]<br>
Line 46: Line 46:
!Lane||Sample||Enzyme1||Enzyme2
!Lane||Sample||Enzyme1||Enzyme2
|-
|-
-
|1||GGA3-1|| || ||  
+
|1||GGA3-1|| ||  
|-
|-
-
|2||GGA3-2|| || ||  
+
|2||GGA3-2|| ||  
|-
|-
-
|3||GGA3-3|| || ||  
+
|3||GGA3-3|| ||  
|-
|-
-
|4||1kbp ladder|| || ||  
+
|4||1kbp ladder|| ||  
|-
|-
-
|5||GGA3-4|| || ||  
+
|5||GGA3-4|| ||  
|-
|-
-
|6||GGA3-5|| || ||  
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|6||GGA3-5|| ||  
|-
|-
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|7||GGA3-6|| || ||  
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|7||GGA3-6|| ||  
|-
|-
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|8||GGA3-7|| || ||  
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|8||GGA3-7|| ||  
|}
|}
[[File:xxxxx.png]]<br>
[[File:xxxxx.png]]<br>
Line 68: Line 68:
!Lane||Sample||Enzyme1||Enzyme2
!Lane||Sample||Enzyme1||Enzyme2
|-
|-
-
|1||GGA3-8|| || ||  
+
|1||GGA3-8|| ||  
|-
|-
-
|2||GGA4-1|| || ||  
+
|2||GGA4-1|| ||  
|-
|-
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|3||GGA4-2|| || ||  
+
|3||GGA4-2|| ||  
|-
|-
-
|4||GGA4-3|| || ||  
+
|4||GGA4-3|| ||  
|-
|-
-
|5||1kbp ladder|| || ||  
+
|5||1kbp ladder|| ||  
|-
|-
-
|6||GGA4-4|| || ||  
+
|6||GGA4-4|| ||  
|-
|-
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|7||GGA4-5|| || ||  
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|7||GGA4-5|| ||  
|-
|-
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|8||GGA4-6|| || ||  
+
|8||GGA4-6|| ||  
|}
|}
[[File:xxxxx.png]]<br>
[[File:xxxxx.png]]<br>
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!Lane||Sample||Enzyme1||Enzyme2
!Lane||Sample||Enzyme1||Enzyme2
|-
|-
-
|1||GGA4-7|| || ||  
+
|1||GGA4-7|| ||  
|-
|-
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|2||GGA4-8|| || ||  
+
|2||GGA4-8|| ||  
|-
|-
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|3||1kbp ladder|| || ||  
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|3||1kbp ladder|| ||  
|-
|-
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|4||GGA7-1|| || ||  
+
|4||GGA7-1|| ||  
|-
|-
-
|5||GGA16-1|| || ||  
+
|5||GGA16-1|| ||  
|-
|-
-
|6||NC(RBS-GFP-DT)|| || ||  
+
|6||NC(RBS-GFP-DT)|| ||  
|}
|}
[[File:xxxxx.png]]<br>
[[File:xxxxx.png]]<br>

Revision as of 13:43, 26 September 2013

Contents

Sep 26

Electrophoresis

No name

LaneSampleEnzyme1Enzyme2
1GGA1-1
2GGA1-2
3GGA1-3
4GGA1-4
5GGA1-5
6GGA1-6
7GGA1-7
8GGA1-8
91kbp ladder
10GGA2-1
11GGA2-2
12GGA2-3
13GGA2-4
14GGA2-5
15GGA2-6
16GGA2-7
17GGA2-8

File:Xxxxx.png

LaneSampleEnzyme1Enzyme2
1GGA3-1
2GGA3-2
3GGA3-3
41kbp ladder
5GGA3-4
6GGA3-5
7GGA3-6
8GGA3-7

File:Xxxxx.png
</div>

LaneSampleEnzyme1Enzyme2
1GGA3-8
2GGA4-1
3GGA4-2
4GGA4-3
51kbp ladder
6GGA4-4
7GGA4-5
8GGA4-6

File:Xxxxx.png
</div>

LaneSampleEnzyme1Enzyme2
1GGA4-7
2GGA4-8
31kbp ladder
4GGA7-1
5GGA16-1
6NC(RBS-GFP-DT)

File:Xxxxx.png
</div>

Ligasion(Golden Gate Assenbly)

No name

stateVectorInserter1Inserter2Inserter3Inserter4Inserter5Ligation High ver.2
experimentRBS-GFP-DT1.0 µLPcon-pT181 antisense(E-2A)0.6µLDT(2-1)0.5µLPcon-pT181 attenuator(1-SA)2.1 µL--------2.1 µL
experimentpSB1C3(EcoRI&SpeI)1.8 µLPcon-pT181 antisense(E-2A)0.9 µLDT(2-1)0.5 µLPcon-GFP-DT(1-SA)1.5 µL--------2.5 µL
experimentRBS-GFP-DT1.0 µLPtet(E-0)0.7µLPcon pT181 attenuator(0-1A)0.3µLtet aptamer(1-3)0.3 µLPcon-tetR-DT(3-2A)1.1 µLPtet(2-5)0.3 µL2.2 µL
experimentRBS-GFP-DT1.0 µLPtet(E-0)0.7 µLPcon-pT181 attenuator-DT(0-1A)1.0 µLPcon-tetR-DT(1-2A)1.0 µLPtet(2-5)0.3µL----2.0 µL
experimentRBS-GFP-DT1.0 µLPcon-pT181 attenuator-tetR-DT(E-1A)1.0 µLPcon antisense-spinach-DT(1-2A)0.7µLPcon-pT 181 attenuator(2-SA)2.1 µL--------2.4 µL
experimentRBS-GFP-DT1.0 µLPcon antisense(E-1A)0.5 µLspinach-DT(1-2)1.0 µLPcon-GFP-DT(1-2)0.8 µL--------2.15 µL
experimentpSB1C3(EcoRI&SpeI)1.8 µLPcon-pT181 antisense(E-1A)0.8µLspinach-DT(1-2)1.0µLPcon-GFP-DT(2-SA)0.8 µL--------2.1 µL
experimentpSB1C3(EcoRI&SpeI)1.8 µLPcon-tet aptamer-DT(E-1A)3.0 µLPtet-pT181 antisense(1-3)1.5 µLspinach-DT(3-2)1.5 µLPcon-GFP-DT(2-5A)0.8 µL----4.3 µL

Liquid Culture

No name

Samplemedium
Pcon-GFP-DTAmp
RBS-GFP-DTCP
Pcon-pT181 attenuator-DTAmp
Pcon-tet aptamer-DTAmp
Pcon-spinach-DTAmp
Pcon-pT181 antisense-spinach-DTAmp
spinach-DTCP

BSAI Digestion

No name

DNAMilliQBSAI-HF10*cut smarttotal
Pcon7 µL10.7 µL0.3 µL2 µL20 µL
RBS-luxR-DT10 µL7 µL0.3 µL2 µL20 µL

File:Xxxxx.jpg

Colony PCR

Tatsui

Samplebase pair
GGA-1 (9~16)3177
GGA-2 (9~16)3189
GGA-3 (9~21)2989
GGA-4 (9~16)2939
GGA-6 (1)3359
GGA-16(2~4)2939
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s3min30s30cycles

Electrophoresis

No name

LaneSampleEnzyme1Enzyme2
1GGA-1 (9)----
2GGA-1 (10)----
3GGA-1 (11)----
4GGA-1 (12)----
5GGA-1 (13)----
6GGA-1 (14)----
7GGA-1 (15)----
8GGA-1 (16)----
91kbp ladder----
10GGA-2 (9)----
11GGA-2 (10)----
12GGA-2 (11)----
13GGA-2 (12)----
14GGA-2 (13)----
15GGA-2 (14)----
16GGA-2 (15)----
17GGA-2 (16)----

File:Xxxxx1.jpg

LaneSampleEnzyme1Enzyme2
1GGA-4 (9)----
2GGA-4 (10)----
3GGA-4 (11)----
4GGA-4 (12)----
5GGA-4 (13)----
6GGA-4 (14)----
7GGA-4 (15)----
8GGA-4 (16)----
91kbp ladder----
10GGA-6 (1)----
11GGA-16 (2)----
12GGA-16 (3)----
13GGA-16 (4)----

File:Xxxxx1.jpg
</div>

|1||GGA-3 (9)||--||-- |- |2||GGA-3 (10)||--||-- |- |3||GGA-3 (11)||--||-- |- |4||GGA-3 (12)||--||-- |- |5||1kbp ladder||--||-- |- |6||GGA-3 (13)||--||-- |- |7||GGA-3 (15)||--||-- |- |8||GGA-3 (14)||--||-- |} File:Xxxxx1.jpg
</div>

|1||GGA-3 (17)||--||-- |- |2||GGA-3 (18)||--||-- |- |3||GGA-3 (19)||--||-- |- |4||1kbp ladder||--||-- |- |5||GGA-3 (20)||--||-- |- |6||GGA-3 (21)||--||-- |- |7||GGA-3 (16)||--||-- |} File:Xxxxx1.jpg
</div>

Electrophoresis

No name

LaneSampleEnzyme1Enzyme2
1RBS-GFP-DT(RNA)----
21kbp ladder----
3Pcon-RBS-GFP-DT(RNA)----

File:Xxxxx1.jpg

Liquid Culture

Hirano

Samplemedium
Pcon-GFP-DT--
RBS-GFP-DT--
Pcon-tet aptamer-DT--
Pcon-spinach-DT--
Pcon-pT181 antisense-spinach-DT--
spinach-DT--

Colony PCR

Hirano

Samplebase pair
9/25 Pcon-pH181 attenuator+RBS-GFP-DT-1--
9/25 Pcon-pH181 attenuator+RBS-GFP-DT-2--
9/25 Pcon-pH181 attenuator+RBS-GFP-DT-3--
9/25 Pcon-pH181 attenuator+RBS-GFP-DT-4--
9/24 Ptet-RBS-GFP-DT-1--
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s1min40s30cycles
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