"
Team:Chiba/Calendar-Home/25 September 2008
From 2008.igem.org
24 September 2008 <|> 26 September 2008
Laboratory work
Team:Communication
Team:Output
colony PCR
Vector:BBa_R0062 Insert:BBa_E0240
Vector:BBa_R0062
| Sample No | each |
| culture | 1 |
| Fwd primer | 1.5 |
| Rev primer | 1.5 |
| Taq(DNA pol) | 0.2 |
| dNTPmix | 3 |
| thermo pol Buffer | 3 |
| dH2O | 19.8 |
| TOTAL | 30 |
->95℃ 5min ->(95℃ 1min 50℃ 30sec 72℃ 1min)x25 ->72℃ 10min
Digestion Test
BBa_I6206
BBa_I6101
BBa_I6042
BBa_I0462
| Sample No | each |
| DNA | 2 |
| EcoRⅠ | 0.2 |
| EcoRⅠ Buffer | 1 |
| dH2O | 7 |
| TOTAL | 30 |
->30min 37℃
Gel cheack
BBa_I6206(digestion product)
BBa_I6101(digestion product)
BBa_I6042(digestion product)
BBa_I0462(digestion product)
BBa_I0642(natural)
| Sample No | each(digestion) | I0462(natural) |
| DNA | 10 | 1 |
| Loading Dye | 2 | 1 |
| dH2O | - | 4 |
| TOTAL | 12 | 6 |
Digestion
BBa_I0462
| DNA | 25 |
| XbaⅠ | 1 |
| PstⅠ | 1 |
| Buffer 3 | 4 |
| BSA | 4 |
| dH2O | 5 |
| TOTAL | 40 |
->3hour 37℃
->Zymo ->10μl
Ligation
Vector:BBa_I7100
Insert:BBa_R0062
Insert:BBa_I0462
| I7100 | 1.3 | 1.3 |
| R0062 | 2 | - |
| I0462 | 3.3 | - |
| Ligase | 1 | 1 |
| Ligase Buffer | 2 | 2 |
| dH2O | 0.4 | 5 |
| TOTAL | 10 | 10 |
->3hour 37℃
->Transformation
