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Template:Kyoto/Notebook/Aug 11
From 2013.igem.org
Contents |
Aug 11
Miniprep
| 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
| 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
| 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
| 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 |
