From 2013.igem.org
Aug 11
Miniprep
Nakamoto
| DNA | concentration[µg/mL]
|
| 8/9 BBa_J44000 -(2) | 100
|
| 8/9 BBa_J23106 | 154
|
| 8/9 BBa_J23100 | 144
|
| 8/10 T7-His-FT | 92
|
Colony PCR
Kojima and Yasuda
| Sample
|
| BBa_B0034 -(2)
|
| BBa_B0034 -(1)
|
| BBa_R0011 -(2)
|
| BBa_R0011 -(1)
|
| negative control
|
| PreDenature | Denature | Annealing | Extension | cycle
|
| 94 °C | 94 °C | 55 °C | 68 °C | --
|
| 5 min | 30 s | 30 s | 20 s | 30 cycles
|
| Sample
|
| BBa_D0412 -(1)
|
| BBa_D0412 -(2)
|
| negative control
|
| PreDenature | Denature | Annealing | Extension | cycle
|
| 94 °C | 94 °C | 55 °C | 68 °C | --
|
| 5 min | 30 s | 30 s | 1 min 30 s | 30 cycles
|
Restriction Enzyme Digestion
Kojima anda Yasuda
| 8/9 BBa_J44000 2 | EcoRI | PstI | 10x buffer H | MilliQ | total
|
| 2 | 0.2 | 0.2 | 1 | 6.6 | 10
|
| 2 | 0.2 | -- | 1 | 6.8 | 10
|
| 2 | -- | 0.2 | 1 | 6.8 | 10
|
| 2 | -- | -- | 1 | 7.0 | 10
|
| 8/9 BBa_J23106 | EcoRI | PstI | 10x buffer H | MilliQ | total
|
| 1.3 | 0.2 | 0.2 | 1 | 7.3 | 10
|
| 1.3 | 0.2 | -- | 1 | 7.5 | 10
|
| 1.3 | -- | 0.2 | 1 | 7.5 | 10
|
| 1.3 | -- | -- | 1 | 7.7 | 10
|
| 8/9 BBa_J23100 2 | EcoRI | PstI | 10x buffer H | MilliQ | total
|
| 1.3 | 0.2 | 0.2 | 1 | 7.3 | 10
|
| 1.3 | 0.2 | -- | 1 | 7.5 | 10
|
| 1.3 | -- | 0.2 | 1 | 7.5 | 10
|
| 1.3 | -- | -- | 1 | 7.7 | 10
|
- incubated at 37 °C for 1 hour.
Electrophoresis
No name
| Lane | Sample | Enzyme1 | Enzyme2
|
| 1 | 100bp ladder | -- | --
|
| 2 | 8/9 BBa_J44000 | EcoRI | PstI
|
| 3 | 8/9 BBa_J44000 | EcoRI | --
|
| 4 | 8/9 BBa_J44000 | -- | PstI
|
| 5 | 8/9 BBa_J44000 | -- | --
|
| 6 | 8/9 BBa_J23106 | EcoRI | PstI
|
| 7 | 8/9 BBa_J23106 | EcoRI | --
|
| 8 | 8/9 BBa_J23106 | -- | PstIs
|
| 9 | 8/9 BBa_J23106 | -- | --
|
| 10 | 8/9 BBa_J23100 | EcoRI | PstI
|
| 11 | 8/9 BBa_J23100 | EcoRI | --
|
| 12 | 8/9 BBa_J23100 | -- | PstI
|
| 13 | 8/9 BBa_J23100 | -- | --
|
| Lane | Sample
|
| 1 | 100bp ladder
|
| 2 | BBa_B0034 (1)
|
| 3 | BBa_B0034 (2)
|
| 4 | BBa_R0011 (1)
|
| 5 | BBa_R0011 (2)
|
| 6 | NC
|
| 7 | 100bp ladder
|
| Lane | Sample
|
| 1 | 1kb ladder
|
| 2 | BBa_P0412 (1)
|
| 3 | BBa_P0412 (2)
|
| 4 | NC
|
File:Aug11electrophoresis3.jpg
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