NYMU Taipei/Part Assembly

From 2007.igem.org

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Line 2: Line 2:
**Restriction enzymes: 1ul each
**Restriction enzymes: 1ul each
**Incubate for 2h
**Incubate for 2h
-
**Heat inactivation in 65°C, 20min
+
**Heat inactivation at 65°C, 20min
-
* Gel Separation (1.5h)
+
* 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