From 2007.igem.org

Revision as of 10:05, 1 September 2007 (view source) Lihsiangyen (Talk | contribs) ← Older edit		Revision as of 10:10, 1 September 2007 (view source) Lihsiangyen (Talk | contribs) Newer edit →
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	**Incubate for 2h		**Incubate for 2h
	**Heat inactivation at 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

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Revision as of 10:10, 1 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 (16h)
  • DNA: 2ul
  • Competent cell: 50ul
• Liquid Culture (16h)
  • 5ml LB
• Plasmid Extraction (1hr)
  • Qiagene Miniprep Kit
• Enzyme digestion to ligation formula