"
Team:Chiba/jk/γ/week 3
From 2008.igem.org
Contents |
WEEK 3
31,August,2008
- BBa_R0010①
- PCR product(BBa_J52008)②
| Sample No. | ① | ② |
| DNA | 10 | 10 |
| SpeⅠ | 1 | 0 |
| PstⅠ | 1 | 1 |
| XbaⅠ | 0 | 1 |
| BSA(x100) | 0.5 | 0.5 |
| Buffer2 | 5 | 0 |
| Buffer3 | 0 | 5 |
| dH2O | 28.5 | 28.5 |
| TOTAL | 50 | 50 |
-->37℃ 3hour
-->①20μl refine
-->②20μl refine
1,September,2008
- vector:BBa_R0010 insert:BBa_J52008①
- negative control:BBa_R0010②
| Sample No. | ① | ② |
| vector | 2 | 2 |
| insert | 3 | 0 |
| Ligase Buffer | 2 | 2 |
| Ligase | 1 | 1 |
| dH2O | 3 | 5 |
| TOTAL | 10 | 10 |
-->R/T 2hour
- BBa_R0079(Las promoter)
- BBa_R0071(RhlR promoter)
- BBa_R0077(cinR promoter+RBS)
- BBa_R0078(cinR promoter)
- BBa_R0062(Lux promoter)
2,September,2008
Colony PCR
| Sample No. | ① |
| culture | 1 |
| Fwd primer | 1.5 |
| Rev primer | 1.5 |
| Thermo pol Buffer | 3 |
| dNTP mix | 3 |
| Taq DNA pol (NEB) | 0.2 (1 unit) |
| dH2O | 19.8 |
| TOTAL | 30 |
3,September,2008
| Sample No. | ①~⑤ |
| DNA tamplate | 5 |
| Buffer3 | 1 |
| dH2O | 4 |
| EcoRⅠ | 0.1 |
| TOTAL | 10 |
-->37℃30min
-->Gel check
| Sample No. | ①~⑤ |
| DNA(digestion sample) | 10 |
| loading Dye | 2 |
| TOTAL | 12 |
4,Septempber,2008
- Ligation product
- rLUciferase(BBa_J52008+BBa_F2620 ) x3
- Venus YFP(BBa_E2030+BBa_F2620) x3
5,September,2008
| DNA(digested sample) | 2 |
| PstⅠ | 0.1 |
| BSA | 0.8 |
| Buffer3 | 1 |
| dH2O | 7 |
| TOTAL | 10 |
-->37℃ 30in
- BBa_J52008+BBa_F2620 x2①
- BBa_E2030+BBa_F2620 x2②
- BBa_J52008(PCR Product)③
- BBa_E2050④
| Sample No. | ①~② | ③,④ |
| DNA(digested sample) | 10 | 1 |
| loading Dye | 2 | 1 |
| TOTAL | 12 | 6 |
- Luciferase check-->no light
| ホーム | メンバー紹介 | プロジェクト紹介 | Parts Submitted to the Registry | モデリング | ノート |
|---|
