Template:Kyoto/Notebook/Sep 27

From 2013.igem.org

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Revision as of 06:35, 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

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

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

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

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

Electrophoresis

No name

LaneSampleEnzyme1Enzyme2
1pSB4K5EP
2pSB4K5--
31kbp ladder--
4Pcon-attenuatorES
5Pcon-attenuator--

File:Igku xxxxxx.xxx

LaneSampleEnzyme1Enzyme2
1RBS-GFP-DTXP
2RBS-GFP-DT--
3RBS-GFP-DTEX
41kbp ladder--
5PtetES
6Ptet--

File:Igku xxxxxx.xxx

Gel Extraction

No name

LaneDNAEnzyme
1100bp ladder--
3PtetE+S
4PtetE+S
5PtetE+S

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

LaneDNAEnzyme
11kbp ladder--
3Pcon-antisense-aptamer-DTX+P
4Pcon-antisense-aptamer-DTX+P
5Pcon-antisense-aptamer-DTX+P

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

LaneDNAEnzyme
11kbp ladder--
3RBS-GFP-DTE+X
4RBS-GFP-DTE+X
5RBS-GFP-DTE+X

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

LaneDNAEnzyme
11kb ladder--
2Pcon-attenuatorE+S
3Pcon-attenuatorE+S
4Pcon-attenuatorE+S
6pSB4K5E+P
7pSB4K5E+P
8pSB4K5E+P
10RBS-GFP-DTX+P
11RBS-GFP-DTX+P
12RBS-GFP-DTX+P

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 (E+S) 10.0ng/µL10µL9/26 pT181 attenuator(E+S) 7.5ng/µL1.3µL3.5 µL
experiment9/25 DT (E+X) 42.8ng/µL2.3µL9/22 Pcon-pT181 antisense(E+S) 46.7ng/µL1.5µL1.9 µL
experiment9/27 pSB4K5(E+P) 12.9ng/µL7.8µL9/10 Pcon-attenuator(E+S) 13.8ng/µL8.0µL3.5 µL
experiment9/27 pSB4K5(E+P) 12.9ng/µL7.8µL9/25 RBS-GFP-DT(X+P) 13.9ng/µL12µL3.5 µL
experiment9/8 pSB4K5 (E+S) 17.6ng/µLxx µL8/21 Pcon-GFP-DT(E+S) 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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