NYMU Taipei/Part Assembly
From 2007.igem.org
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**Restriction enzymes: 1ul each | **Restriction enzymes: 1ul each | ||
**Incubate for 2h | **Incubate for 2h | ||
| - | **Heat inactivation | + | **Heat inactivation at 65°C, 20min |
| - | * Gel Separation ( | + | * Gel Separation (0.5h) |
**100V, 30min | **100V, 30min | ||
| + | * Gel Extraction (0.5h) | ||
| + | **Qiagene GelExtraction Kit | ||
* Ligation (3hr) | * Ligation (3hr) | ||
**T4 ligase: 1ul | **T4 ligase: 1ul | ||
**Incubate at RT for 3h | **Incubate at RT for 3h | ||
| - | * Plate Culture (16h) | + | * Plate Culture and Transformation(16h) |
**DNA: 2ul | **DNA: 2ul | ||
**Competent cell: 50ul | **Competent cell: 50ul | ||
| Line 15: | Line 17: | ||
* Plasmid Extraction (1hr) | * Plasmid Extraction (1hr) | ||
**Qiagene Miniprep Kit | **Qiagene Miniprep Kit | ||
| + | * Enzyme digestion to ligation formula | ||
| + | ** [[NYMU Taipei/phptest | phptest]] | ||
Latest revision as of 05:56, 16 September 2007
- Enzyme Digestion (2h)
- Restriction enzymes: 1ul each
- Incubate for 2h
- Heat inactivation at 65°C, 20min
- Gel Separation (0.5h)
- 100V, 30min
- Gel Extraction (0.5h)
- Qiagene GelExtraction Kit
- Ligation (3hr)
- T4 ligase: 1ul
- Incubate at RT for 3h
- Plate Culture and Transformation(16h)
- DNA: 2ul
- Competent cell: 50ul
- Liquid Culture (16h)
- 5ml LB
- Plasmid Extraction (1hr)
- Qiagene Miniprep Kit
- Enzyme digestion to ligation formula
