From 2013.igem.org
Sep 23
No Name
| Lane | DNA | Enzyme
|
| 1 | 1kbp ladder |
|
| 3-4 | Pcon-RBS-tetR-PT |
|
File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx
| Name | concentration[µg/mL] | 260/280 | 260/230
|
| Pcon-PT181 anti E+S | 46.7 | 1.06 | 0.93
|
| Pcon-RBS-tetR-DT E+X | 119.3 | 0.99 |
|
| RBS-8FP-DT E+X | 87.7 | 1.00 | 0.98
|
| ap42_1R-DT E+X | 68.8 | 1.32 | 1.03
|
| Pcon-RBS-tetR-DT 1-2 A | 203.0 | 1.66 | 1.79
|
| Pcon-RBS-tetR-DT 3-2 A | 67.7 | 2.64 | 1.28
|
| Pcon-RBS-GFP-DT 1-S A | 185.6 | 1.88 | 2.04
|
| Pcon-RBS-GFP-DT 2-S A | 298.6 | 1.68 | 1.55
|
| Pcon-tetR-DT E-2 A | 202.4 | 1.57 | 1.66
|
PCR
No name
| genome DNA | KOD plus | 10x buffer | dNTP | MgSO4 | SasA_fwd primer | SasA_rev primer | MilliQ | total
|
| 9/8 Pcon-pT181 anti | 0.5 | 2.5 | 2.5 | 1.5 | 0.75 | 0.75 | 15.5 | 25
|
| 8/20 Ptet(1) | 0.5 | 2.5 | 2.5 | 1.5 | 0.75 | 0.75 | 15.5 | 25
|
| 9/16 Plac 2 | 0.5 | 2.5 | 2.5 | 1.5 | 0.75 | 0.75 | 15.5 | 25
|
| PreDenature | Denature | Annealing | Extension | cycle
|
| 94 °C | 98 °C | 57 °C | 68 °C | --
|
| 2min | 10sec | 30sec | 20sec | 30cycles
|
Ligation
Yoshida
| state | Vector | Inserter | Ligation High ver.2
|
| experiment | 9/21 Pcon-RBS-GFP-DT E+X | 47.5 µg/mL | 9/15 Pcon-RBS-tetR-DT E+S | 59 µg/mL | 2.3 µg/mL
|
| experiment | 9/21 Pcon-RBS-GFP-DT E+X | 47.5 µg/mL | 9/15 Pcon-apt12_1R-DT E+S | 17 µg/mL | 2.4 µg/mL
|
| experiment | 9/21 Pcon-RBS-GFRDT E+X | 2.1 µg/mL | 9/15 Pcon-DT181 atte-DT | 21 µg/mL | 3.0 µg/mL
|
| experiment | 9/22 apt 12_1R-DT E+X | 67.8 µg/mL | 9/18 Pcon-DT181 atte E+S | 5.7 µg/mL | 3.5 µg/mL
|
| experiment | 9/22RBS-GFP-DT E+X | 87.7 µg/mL | 9/18 Pcon-DT18 atte E+S | 6.7 µg/mL | 3.5 µg/mL
|
| experiment | 9/13 DT E+X | 25.4 µg/mL | 9/22 Pcon-DT181 anti E+S | 46.8 µg/mL | 2.3 µg/mL
|
PCR
No name
| genome DNA | KOD plus | 10x buffer | dNTP | MgSO4 | SasA_fwd primer | SasA_rev primer | MilliQ | total
|
| Pcon-atte-DT (E-1A) | 0.5 | 2.5 | 2.5 | 1.5 | 0.75 | 0.75 | 77.5 | 25
|
| Pcon-atte-DT (2-1A) | 0.5 | 2.5 | 2.5 | 1.5 | 0.75 | 0.75 | 77.5 | 25
|
| Pcon-anti-spin-DT (1-2A) | 0.5 | 2.5 | 2.5 | 1.5 | 0.75 | 0.75 | 77.5 | 25
|
| Pcon-anti-spin-DT (E-1A) | 0.5 | 2.5 | 2.5 | 1.5 | 0.75 | 0.75 | 77.5 | 25
|
| PreDenature | Denature | Annealing | Extension | cycle
|
| 94 °C | 98 °C | 57 °C | 68 °C | --
|
| 2min | 10s | 30s | 45s | 30cycle
|
Miniprep
No name
| DNA | concentration[µg/mL] | 260/280 | 260/230
|
| 9/22 Plac-DT181 atte-1 | 204.0 | 2.00 | 1.95
|
| 9/22 Plac-DT181 atte-2 | 195.3 | 1.84 | 1.62
|
| 9/22 Plac-DT181 anti-1 | 188.2 | 1.87 | 2.15
|
| 9/22 Plac-DT181 anti-2 | 165.0 | 1.95 | 2.66
|
Electrophoresis
Hirano
| Lane | Sample | Enzyme1 | Enzyme2
|
| 1,6 | 100bp ladder(Promega) | |
|
| 2 | Pcon-atte-DT (E-1A) | |
|
| 3 | Pcon-atte-DT (2-1A) | |
|
| 4 | Pcon-anti-spi-DT (1-2A) | |
|
| 5 | Pcon-anti-spi-DT (E-1A) | |
|
File:Igku xxxxxx.xxx
Hirano
| Name | concentration[µg/mL] | 260/280 | 260/230
|
| Pcon atte-DT(E-1A) | 166.7 | 1.51 | 0.99
|
| Pcon atte-DT(2-1A) | 122.1 | 1.75 | 1.20
|
| Pcon anti spi DT(1-2A) | 99.1 | 1.86 | 1.81
|
Electrophoresis
Hirano
| Lane | Sample | Enzyme1 | Enzyme2
|
| 100bp ladder | | |
|
| 9/8 Pcon-pT181 anti(E-1A) | | |
|
| 8/20 Ptet(1)(2-S) | | |
|
| 9/16 Plac2(E-1A) | | |
|
File:Igku xxxxxx.xxx
Transformation
No Name
| Name | Sample | Competent Cells | Total | Plate
|
| pSB1C3 | 2µL | 20µL | 22µL |
|
Ligation
No Name
| state | Vector | Inserter | Ligation High ver.2
|
| experiment | 9/19 Plac S+P | 24.6 µg/mL | 9/18 ape12_1R-DT X+P | 4.5 µg/mL | 2 µg/mL
|
| experiment | 9/21 RBS-GFP-DT E+X | 88.4 µg/mL | 9/15 Pcon-pT181 atte E+S | 21 µg/mL | 2 µg/mL
|
| experiment | 9/13 PSB1C3 X+P | 26.0 µg/mL | 9/16 pT181 atte X+P | 8.2 µg/mL | 2 µg/mL
|
"
