NYMU Taipei/Part Assembly

From 2007.igem.org

(Difference between revisions)
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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