NYMU Taipei/Lab Notes/2007 8 27
From 2007.igem.org
< NYMU Taipei/Lab Notes(Difference between revisions)
(→plasmid concentration after extraction) |
(→plasmid concentration after extraction) |
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| Line 6: | Line 6: | ||
|- | |- | ||
|pOmpC | |pOmpC | ||
| - | | | + | |0.06 |
|0.006 | |0.006 | ||
|- | |- | ||
|EYFP | |EYFP | ||
| - | | | + | |0.3 |
|0.030 | |0.030 | ||
|- | |- | ||
|ECFP | |ECFP | ||
| - | |23 | + | |0.23 |
|0.023 | |0.023 | ||
|- | |- | ||
|CinR | |CinR | ||
| - | |15 | + | |0.15 |
|0.015 | |0.015 | ||
|- | |- | ||
|HSL | |HSL | ||
| - | |49 | + | |0.49 |
|0.049 | |0.049 | ||
|- | |- | ||
|pCinHSL | |pCinHSL | ||
| - | | | + | |0.07 |
|0.007 | |0.007 | ||
|- | |- | ||
|pCinHSLRBS | |pCinHSLRBS | ||
| - | |12 | + | |0.12 |
|0.012 | |0.012 | ||
|- | |- | ||
|RBS | |RBS | ||
| - | |14 | + | |0.14 |
|0.014 | |0.014 | ||
|- | |- | ||
|D-term | |D-term | ||
| - | |27 | + | |0.27 |
|0.027 | |0.027 | ||
|- | |- | ||
Latest revision as of 06:19, 16 September 2007
Contents |
plasmid concentration after extraction
| part | concentration (ug/ul) | A260 (OD) |
|---|---|---|
| pOmpC | 0.06 | 0.006 |
| EYFP | 0.3 | 0.030 |
| ECFP | 0.23 | 0.023 |
| CinR | 0.15 | 0.015 |
| HSL | 0.49 | 0.049 |
| pCinHSL | 0.07 | 0.007 |
| pCinHSLRBS | 0.12 | 0.012 |
| RBS | 0.14 | 0.014 |
| D-term | 0.27 | 0.027 |
- p.s.1: the sample is diluted to 200X, and one OD is 50 ug/mL
- p.s.2: the calculation of concentration should be OD multiplied only by 10, not by 1,000 (It is a mistake)
- p.s.3: 50 ug/mL = 0.05 ug/uL, and sample is diluted to 200X. Thus the original concentration should be 0.05 ug/uL * 200 = 10 ug/uL for per OD
- p.s.4: Eppendorf®Biophotometer
Phase 1 assembly
- Objective: pOmpC + (RBS+EYFP)
- 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)
- pOmpC: 14ul
- SpeI: 1ul
- PstI: 1ul
- 10x buffer: 2, 2ul
- 10x BSA: 2ul
- Total: 20ul
- 37°C, 2h
- Heat inactivation: 65°C 20min
- Enzyme Digestion (EYFP)
- EYFP: 3ul
- XbaI: 1ul
- PstI: 1ul
- 10x buffer: 3, 2ul
- 10x BSA: 2ul
- H2O: 11ul
- Total: 20ul
- 37°C, 2h
- Heat inactivation: 65°C 20min
Others
- liquid culture for tar-EnvZ, pLacIQ in 5 ml LB (Kan:20ug/ml)
Who attend?
- 立翔
- 德銘
- 惇茹
- 香君
- 宥嫺
