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August/20 August 2009
From 2009.igem.org
(Difference between revisions)
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↓<br> | ↓<br> | ||
gel electrophoresis<br> | gel electrophoresis<br> | ||
| + | [[Image:Osaka090819.jpg]]<br> | ||
| + | UP : 1-2M , 1-14F , 1-2M , 2-8O | ||
| + | DOWN : No.3-A,B,C,D , No.4-A,B,C,D , No.6-A,B,C,D , Vector | ||
gel cut<br> | gel cut<br> | ||
↓<br> | ↓<br> | ||
| Line 57: | Line 60: | ||
⑥(RBS+cinI),K204006<br> | ⑥(RBS+cinI),K204006<br> | ||
⑥-A,B,C : OK | ⑥-A,B,C : OK | ||
| + | |||
| + | |||
| + | [https://2009.igem.org/Team:Osaka/NOTES back to NOTES] | ||
Latest revision as of 08:00, 12 October 2009
1.digestion and ligation
digestion with restriction enzyme
K204007 , (K204003)
| Vector | Insert | ||
|---|---|---|---|
| RBS([http://partsregistry.org/wiki/index.php?title=Part:BBa_B0034 1-2M]) | 10 | cinR([http://partsregistry.org/wiki/index.php?title=Part:BBa_C0077 1-14F]) | 12 |
| SpeI | 1 | XbaI | 1 |
| PstI | 1 | PstI | 1 |
| No.2 Buffer | 2 | No.2 | 2 |
| dH2O | 6 | dH2O | 4 |
| total | 20uL | total | 20uL |
K204008
| Vector | Insert | ||
|---|---|---|---|
| RBS([http://partsregistry.org/wiki/index.php?title=Part:BBa_B0034 1-2M]) | 10 | LuxI([http://partsregistry.org/wiki/index.php?title=Part:BBa_C0161 2-8O]) | 12 |
| SpeI | 1 | XbaI | 1 |
| PstI | 1 | PstI | 1 |
| No.2 Buffer | 2 | No.2 | 2 |
| dH2O | 6 | dH2O | 4 |
| total | 20uL | total | 20uL |
↓
37°C, 2hr
↓
gel electrophoresis
UP : 1-2M , 1-14F , 1-2M , 2-8O DOWN : No.3-A,B,C,D , No.4-A,B,C,D , No.6-A,B,C,D , Vector
gel cut
↓
purification by [QIAquick Nucleotide Removal Kit]
↓
ligation
ligation DNA 44 10* buffer 5 ligation 1 total 50uL ↓ 16°C overnight
2. rapid check
③(RBS+cinR),K204003
③-C,D : OK
④(RBS+LasR),K204004
④-A,B,C : OK
⑥(RBS+cinI),K204006
⑥-A,B,C : OK
