Template:Kyoto/Notebook/Sep 27

From 2013.igem.org

(Difference between revisions)
(→Gel Extraction)
(→Gel Extraction)
Line 106: Line 106:
|1||100bp ladder||--
|1||100bp ladder||--
|-
|-
-
|3||Ptet||E+S
+
|3||Ptet||EcoRI+SpeI
|-
|-
-
|4||Ptet||E+S
+
|4||Ptet||EcoRI+SpeI
|-
|-
-
|5||Ptet||E+S
+
|5||Ptet||EcoRI+SpeI
|}
|}
[[File:igku_xxbeforexx.xxx]]<br>
[[File:igku_xxbeforexx.xxx]]<br>
Line 119: Line 119:
|1||1kbp ladder||--
|1||1kbp ladder||--
|-
|-
-
|3||Pcon-antisense-aptamer-DT||X+P
+
|3||Pcon-antisense-aptamer-DT||XbaI+PstI
|-
|-
-
|4||Pcon-antisense-aptamer-DT||X+P
+
|4||Pcon-antisense-aptamer-DT||XbaI+PstI
|-
|-
-
|5||Pcon-antisense-aptamer-DT||X+P
+
|5||Pcon-antisense-aptamer-DT||XbaI+PstI
|}
|}
[[File:igku_xxbeforexx.xxx]]<br>
[[File:igku_xxbeforexx.xxx]]<br>
Line 132: Line 132:
|1||1kbp ladder||--
|1||1kbp ladder||--
|-
|-
-
|3||RBS-GFP-DT||E+X
+
|3||RBS-GFP-DT||EcoRI+XbaI
|-
|-
-
|4||RBS-GFP-DT||E+X
+
|4||RBS-GFP-DT||EcoRI+XbaI
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|-
-
|5||RBS-GFP-DT||E+X
+
|5||RBS-GFP-DT||EcoRI+XbaI
|-
|-
|}
|}
Line 146: Line 146:
|1||1kb ladder||--
|1||1kb ladder||--
|-
|-
-
|2||Pcon-attenuator||E+S
+
|2||Pcon-attenuator||EcoRI+SpeI
|-
|-
-
|3||Pcon-attenuator||E+S
+
|3||Pcon-attenuator||EcoRI+SpeI
|-
|-
-
|4||Pcon-attenuator||E+S
+
|4||Pcon-attenuator||EcoRI+SpeI
|-
|-
-
|6||pSB4K5||E+P
+
|6||pSB4K5||EcoRI+PstI
|-
|-
-
|7||pSB4K5||E+P
+
|7||pSB4K5||EcoRI+PstI
|-
|-
-
|8||pSB4K5||E+P
+
|8||pSB4K5||EcoRI+PstI
|-
|-
-
|10||RBS-GFP-DT||X+P
+
|10||RBS-GFP-DT||XbaI+PstI
|-
|-
-
|11||RBS-GFP-DT||X+P
+
|11||RBS-GFP-DT||XbaI+PstI
|-
|-
-
|12||RBS-GFP-DT||X+P
+
|12||RBS-GFP-DT||XbaI+PstI
|}
|}
[[File:igku_xxbeforexx.xxx]]<br>
[[File:igku_xxbeforexx.xxx]]<br>

Revision as of 06:43, 27 September 2013

Contents

Sep 27

Gel Extraction

Kojima

LaneDNAEnzyme
11kbp ladder--
2pSB1C3EcoRI+SpeI
3
4
6Pcon-attenuator-aptmer-DTEcoRI+SpeI
7
8
10Pcon-attenuator-aptmer-DTEcoRI+SpeI
11
12

Igku 9272.jpg
Igku 9273.jpg

Nameconcentration[µg/mL]260/280260/230
Pcon-attenuator-DT(EcoRI+SpeI)6.81.980.43
pT181-attenuator(EcoRI+SpeI)7.51.840.06
pSB1C3(EcoRI+SpeI)17.71.970.71

Gel Extraction

No name

LaneDNAEnzyme
（＞ω・）b

Igku 9274.jpg
Igku 9275.jpg

Nameconcentration[µg/mL]260/280260/230
（´ω｀）
（ΦωΦ^）

Electrophoresis

No name

LaneSampleEnzyme1Enzyme2
1pSB4K5EcoRIPstI
2pSB4K5--
31kbp ladder--
4Pcon-attenuatorEcoRISpeI
5Pcon-attenuator--

File:Igku xxxxxx.xxx

LaneSampleEnzyme1Enzyme2
1RBS-GFP-DTXbaIPstI
2RBS-GFP-DT--
3RBS-GFP-DTEcoRIXbaI
41kbp ladder--
5PtetEcoRISpeI
6Ptet--

File:Igku xxxxxx.xxx

Gel Extraction

No name

LaneDNAEnzyme
1100bp ladder--
3PtetEcoRI+SpeI
4PtetEcoRI+SpeI
5PtetEcoRI+SpeI

File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx

LaneDNAEnzyme
11kbp ladder--
3Pcon-antisense-aptamer-DTXbaI+PstI
4Pcon-antisense-aptamer-DTXbaI+PstI
5Pcon-antisense-aptamer-DTXbaI+PstI

File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx

LaneDNAEnzyme
11kbp ladder--
3RBS-GFP-DTEcoRI+XbaI
4RBS-GFP-DTEcoRI+XbaI
5RBS-GFP-DTEcoRI+XbaI

File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx

LaneDNAEnzyme
11kb ladder--
2Pcon-attenuatorEcoRI+SpeI
3Pcon-attenuatorEcoRI+SpeI
4Pcon-attenuatorEcoRI+SpeI
6pSB4K5EcoRI+PstI
7pSB4K5EcoRI+PstI
8pSB4K5EcoRI+PstI
10RBS-GFP-DTXbaI+PstI
11RBS-GFP-DTXbaI+PstI
12RBS-GFP-DTXbaI+PstI

File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx

Nameconcentration[µg/mL]260/280260/230
（´ω｀）
（ΦωΦ^）

Liquid Culture

No name

Samplemedium
Pcon-GFPAmp
RBS-GFP-DTCP
Pcon-attenuatorAmp
Pcon-attenuator-aptamer-DTAmp
Pcon-tetR-DTAmp

Ligation

No name

stateVectorInserterLigation High ver.2
experiment9/17 pSB1C3 (EcoRI+SpeI) 10.0ng/µL10µL9/26 pT181 attenuator(EcoRI+SpeI) 7.5ng/µL1.3µL3.5 µL
experiment9/25 DT (EcoRI+XbaI) 42.8ng/µL2.3µL9/22 Pcon-pT181 antisense(EcoRI+SpeI) 46.7ng/µL1.5µL1.9 µL
experiment9/27 pSB4K5(EcoRI+PstI) 12.9ng/µL7.8µL9/10 Pcon-attenuator(EcoRI+SpeI) 13.8ng/µL8.0µL3.5 µL
experiment9/27 pSB4K5(EcoRI+PstI) 12.9ng/µL7.8µL9/25 RBS-GFP-DT(XbaI+PstI) 13.9ng/µL12µL3.5 µL
experiment9/8 pSB4K5 (EcoRI+SpeI) 17.6ng/µLxx µL8/21 Pcon-GFP-DT(EcoRI+SpeI) 11ng/µL12.6µLxx µL

Liquid Culture

Hirano

Samplemedium
Pcon-spinach-DTplusgrow Amp
Pcon-tetRaptamer-DTplusgrow Amp　
Pcon-antisense-spinach-DTplusgrow Amp
spinach-DTplusgrow CP

RNA Extraction

Nakamoto

Nameconcentration[µg/mL]260/280260/230
9/26 Pcon-GFP-DT423.01.831.46
9/26 Pcon-GFP-DT K 298.71.821.50

Electrophoresis

No name

LaneSample
19/27 Pcon-GFP-DT(RNA)
21kb ladder
39/27 Pcon-GFP-DT K(RNA)

Igku 9271.jpg

Miniprep

Tatsui

DNA
Pcon-aptamer-DT
Pcon-spinach-DT
Pcon-antisense
Pcon-tetR-DT
Pcon-attenuator-DT
Pcon-antisense-spinach-DT

Transformation

Hirano

Name1Name2Sample1(µL)Sample2(µL)Competent Cells(µL)Total(µL)
Pcon-attenuator+RBS-GFP-DT(4K5)Pcon-attenuator-DT(1A2)213033
Pcon-attenuator+RBS-GFP-DT(4K5)Pcon-antisense-spinach-DT(1A2)213033
Pcon-attenuator+RBS-GFP-DT(4K5)Pcon-tetR aptamer-DT(1A2)213033
Pcon-RBS-GFP-DT(4K5)Pcon-attenuator-DT(1A2)213033
Pcon-RBS-GFP-DT(4K5)Pcon-antisense-spinach-DT(1A2)213033
Pcon-RBS-GFP-DT(4K5)Pcon-tetR aptamer-DT(1A2)213033
Pcon-attenuator+RBS-GFP-DT(4K5)--2--2022
Pcon-RBS-GFP-DT(4K5)--2--2022
attenuator(1C3)--2--2022
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