NYMU Taipei/Part Assembly
From 2007.igem.org
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* Enzyme Digestion (2h) | * Enzyme Digestion (2h) | ||
+ | **Restriction enzymes: 1ul each | ||
+ | **Incubate for 2h | ||
+ | **Heat inactivation in 65°C, 20min | ||
* Gel Separation (1.5h) | * Gel Separation (1.5h) | ||
+ | **100V, 30min | ||
* Ligation (3hr) | * Ligation (3hr) | ||
+ | **T4 ligase: 1ul | ||
+ | **Incubate at RT for 3h | ||
* Plate Culture (16h) | * Plate Culture (16h) | ||
+ | **DNA: 2ul | ||
+ | **Competent cell: 50ul | ||
* Liquid Culture (16h) | * Liquid Culture (16h) | ||
+ | **5ml LB | ||
* Plasmid Extraction (1hr) | * Plasmid Extraction (1hr) | ||
+ | **Qiagene Miniprep Kit |
Revision as of 09:31, 1 September 2007
- Enzyme Digestion (2h)
- Restriction enzymes: 1ul each
- Incubate for 2h
- Heat inactivation in 65°C, 20min
- Gel Separation (1.5h)
- 100V, 30min
- Ligation (3hr)
- T4 ligase: 1ul
- Incubate at RT for 3h
- Plate Culture (16h)
- DNA: 2ul
- Competent cell: 50ul
- Liquid Culture (16h)
- 5ml LB
- Plasmid Extraction (1hr)
- Qiagene Miniprep Kit