NYMU Taipei/Lab Notes/2007 8 27
From 2007.igem.org
(Difference between revisions)
(→first assembly) |
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**RE: 1ul | **RE: 1ul | ||
**Heat inactivation: 65°C 20min | **Heat inactivation: 65°C 20min | ||
+ | ---- | ||
+ | |||
==Others== | ==Others== | ||
*Culture tar-EnvZ, pLacIQ in 5 ml LB liquid medium (Kan:20ug/ml) | *Culture tar-EnvZ, pLacIQ in 5 ml LB liquid medium (Kan:20ug/ml) |
Revision as of 13:20, 28 August 2007
- plasmid concentration after liquid culture extraction
part | concentration (ug/ul) |
---|---|
pOmpC | 6 |
EYFP | 30 |
ECFP | 23 |
CinR | 15 |
HSL | 49 |
pCinHSL | 7 |
pCinHSLRBS | 12 |
RBS | 14 |
D-term | 27 |
first assembly
- Ligation
- Insert:vector=3:1
- Vector: 300ng/ul(2ul pOmpC after enzyme digestion)
- Insert: 100ng/ul(2ul EYFP after enzyme digestion)
- Ligase: 1ul
- Final volume: 10ul
- RT 3h
- Enzyme Digestion
- pOmpC: 14ul EYFP: 3ul
- RE: 1ul
- Heat inactivation: 65°C 20min
Others
- Culture tar-EnvZ, pLacIQ in 5 ml LB liquid medium (Kan:20ug/ml)