NYMU Taipei/Lab Notes/2007 9 16
From 2007.igem.org
(Difference between revisions)
| Line 1: | Line 1: | ||
| + | ==Enzyme Digestion== | ||
*Enzyme digestion: CinR+HSL (with RBS) with EcoRI, SpeI | *Enzyme digestion: CinR+HSL (with RBS) with EcoRI, SpeI | ||
**CinR+HSL (1~4): 14ul | **CinR+HSL (1~4): 14ul | ||
| Line 13: | Line 14: | ||
**H2O: 6ul | **H2O: 6ul | ||
**Total: 20ul | **Total: 20ul | ||
| + | ==Gel separation== | ||
| + | * 1% gel | ||
| + | * TAE 1X | ||
| + | * 30 min, 100v | ||
| + | * use 1Kb ladder | ||
| + | ** CinR+HSL insert size = 1.53 Kb | ||
| + | ** D-term vector size = 3.284 Kb | ||
Revision as of 09:10, 16 September 2007
Enzyme Digestion
- Enzyme digestion: CinR+HSL (with RBS) with EcoRI, SpeI
- CinR+HSL (1~4): 14ul
- 因為濃度未知, 所以用最大體積
- EcoRI: 1ul (20,000 units/ml)
- SpeI: 1ul (10,000 units/ml)
- 10x EcoRI buffer: 2ul
- Total: 20ul
- CinR+HSL (1~4): 14ul
- Enzyme digestion: D-term with EcoRI
- D-term: 11ul (損失因子10, 目標重量300ng)
- 300(ng) * 10 = 3(ug) = 0.27 (ug/uL) * X(uL), X = 3/0.27 = 11.11111111
- EcoRI: 1ul
- 10x EcoRI buffer: 2ul
- H2O: 6ul
- Total: 20ul
- D-term: 11ul (損失因子10, 目標重量300ng)
Gel separation
- 1% gel
- TAE 1X
- 30 min, 100v
- use 1Kb ladder
- CinR+HSL insert size = 1.53 Kb
- D-term vector size = 3.284 Kb
