From 2013.igem.org
Sep 27
Kojima
Lane | DNA | Enzyme
|
1 | 1kbp ladder | --
|
2 | pSB1C3 | EcoRI+SpeI
|
3
|
4
|
6 | Pcon-attenuator-aptmer-DT | EcoRI+SpeI
|
7
|
8
|
10 | Pcon-attenuator-aptmer-DT | EcoRI+SpeI
|
11
|
12
|
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
Name | concentration[µg/mL] | 260/280 | 260/230
|
Pcon-attenuator-DT(EcoRI+SpeI) | 6.8 | 1.98 | 0.43
|
pT181-attenuator(EcoRI+SpeI) | 7.5 | 1.84 | 0.06
|
pSB1C3(EcoRI+SpeI) | 17.7 | 1.97 | 0.71
|
Electrophoresis
Liquid Culture
No name
Sample | medium
|
Pcon-GFP | Amp
|
RBS-GFP-DT | CP
|
Pcon-attenuator | Amp
|
Pcon-attenuator-aptamer-DT | Amp
|
Pcon-tetR-DT | Amp
|
Ligation
じっけんしゃ
state | Vector | Inserter | Ligation High ver.2
|
experiment | 9/17 pSB1C3 (E+S) 10.0ng/µL | 10µL | 9/26 pT181 attenuator(E+S) 7.5ng/µL | 1.3µL | xx µL
|
experiment | 9/25 DT (E+X) 42.8ng/µL | 2.3µL | 9/22 Pcon-pT181 antisense(E+S) 46.7ng/µL | 1.5µL | xx µL
|
experiment | 9/27 pSB4K5(E+P) 12.9ng/µL | 7.8µL | 9/10 Pcon-attenuator(E+S) 13.8ng/µL | 8.0µL | xx µL
|
experiment | 9/27 pSB4K5(E+P) 12.9ng/µL | 7.8µL | 9/25 RBS-GFP-DT(X+P) 13.9ng/µL | 12µL | xx µL
|
experiment | 9/8 pSB4K5 (E+S) 17.6ng/µL | xx µL | 8/21 Pcon-GFP-DT(E+S) 11ng/µL | 12.6µL | xx µL
|
Liquid Culture
No name
Sample | medium
|
Pcon-spinach-DT | plusgrow Amp
|
Pcon-tetRaptamer-DT | plusgrow Amp
|
Pcon-antisense-spinach-DT | plusgrow Amp
|
spinach-DT | plusgrow CP
|
Nakamoto
Name | concentration[µg/mL] | 260/280 | 260/230
|
9/26 Pcon-GFP-DT | 423.0 | 1.83 | 1.46
|
9/26 Pcon-GFP-DT K | 298.7 | 1.82 | 1.50
|
Electrophoresis
No name
Lane | Sample
|
1 | 9/27 Pcon-GFP-DT(RNA)
|
2 | 1kb ladder
|
3 | 9/27 Pcon-GFP-DT K(RNA)
|
File:Igku xxxxxx.xxx
Miniprep
Tatsui
DNA
|
Pcon-aptamer-DT
|
Pcon-spinach-DT
|
Pcon-antisense
|
Pcon-tetR-DT
|
Pcon-attenuator-DT
|
Pcon-antisense-spinach-DT
|
Transformation
Hirano
Name1 | Name2 | Sample1(µL) | Sample2(µL) | Competent Cells(µL) | Total(µL)
|
Pcon-attenuator+RBS-GFP-DT(4K5) | Pcon-attenuator-DT(1A2) | 2 | 1 | 30 | 33
|
Pcon-attenuator+RBS-GFP-DT(4K5) | Pcon-antisense-spinach-DT(1A2) | 2 | 1 | 30 | 33
|
Pcon-attenuator+RBS-GFP-DT(4K5) | Pcon-tetR aptamer-DT(1A2) | 2 | 1 | 30 | 33
|
Pcon-RBS-GFP-DT(4K5) | Pcon-attenuator-DT(1A2) | 2 | 1 | 30 | 33
|
Pcon-RBS-GFP-DT(4K5) | Pcon-antisense-spinach-DT(1A2) | 2 | 1 | 30 | 33
|
Pcon-RBS-GFP-DT(4K5) | Pcon-tetR aptamer-DT(1A2) | 2 | 1 | 30 | 33
|
Pcon-attenuator+RBS-GFP-DT(4K5) | -- | 2 | -- | 20 | 22
|
Pcon-RBS-GFP-DT(4K5) | -- | 2 | -- | 20 | 22
|
attenuator(1C3) | -- | 2 | -- | 20 | 22
|