From 2013.igem.org
Sep 17
Liquid Culture
Tatsui and Hirano
Sample | medium
|
Spi-DT | LB medium(+CP)
|
Pcon-Spi-DT,Pcon-pT181-anti-spi-DT | LB medium(+Amp)
|
PCR
Hirano
Pcon-tet ap-DT(VF) | BigDye | 5x buffer | primer(VF) | MilliQ | total
|
1.3 | 0.5 | 1.75 | 0.5 | 0.45 | 10.5
|
PreDenature | Denature | Annealing | Extension | cycle
|
96°C | 96°C | 50°C | 60°C | --
|
2m | 10s | 5s | 4m | 30cycles
|
Pcon-tet ap-DT(VR) | BigDye | 5x buffer | primer(VR) | MilliQ | total
|
1.3 | 0.5 | 1.75 | 0.5 | 0.45 | 10.5
|
PreDenature | Denature | Annealing | Extension | cycle
|
96°C | 96°C | 50°C | 60°C | --
|
2m | 10s | 5s | 4m | 30cycles
|
Ligation
No Name
state | Vector | Inserter | Ligation High ver.2
|
8/28 | name | xx µg/mL | name | xx µg/mL | xx µg/mL
|
NC | name | xx µg/mL | name | xx µg/mL | xx µg/mL
|
Lane | DNA | Enzyme
|
1 | 1kbpladder | --
|
2 | pSB1C3 | E+S
|
3
|
4
|
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
Name | concentration[µg/mL] | 260/280 | 260/230
|
pSB1C3 | 10.0 | 1.84 | 0.40
|
Lane | DNA | Enzyme
|
1 | 1kbpladder | --
|
2 | pSB4K5 | X+P
|
3
|
4
|
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
Name | concentration[µg/mL] | 260/280 | 260/230
|
pSB4K5 | 7.6 | 1.81 | 0.32
|
Lane | DNA | Enzyme
|
1 | 1kbpladder | --
|
2 | Pcon-RBS-tetR-DT | X+P
|
3
|
4
|
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
Name | concentration[µg/mL] | 260/280 | 260/230
|
Pcon-RBS-tetR-DT | 8.0 | 2.40 | 1.14
|
Lane | DNA | Enzyme
|
1 | 1kbpladder | --
|
2 | pSB1C3 | X+P
|
3
|
4
|
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
Name | concentration[µg/mL] | 260/280 | 260/230
|
pSB1C3 | 9.1 | 2.17 | 0.35
|