Team:NTU-LIHPAO-Taiwan/Basic Part

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• Notes
  • Preparation of Competent Cells (E.coli)
  • DNA Dissolution
  • Agarose Gel Electrophoresis
  • Gel Extraction
  • DNA Ligation
  • Plasmid DNA Extraction Using Alkaline Lysis Method (E.coli)
  • Transformation (E.coli)

DNA Ligation

Materials and Reagents :

1. DNA (insert & vector)
2. Ligation high ver.2

Equipment :

1. Cooling dry bath incubator

Procedure :

1. Table

   Components	Volume (μL)
   Insert	x
   Vector	y
   Ligation High	1
   Total	7

           Molar ratio: insert/vector = 3/1
           x + y = 6
   
2. Gently mix the solution by pipetting up and down
3. Incubate
   1. at 16℃ for 2 hours, or
   2. at 37℃ for 1 hour, or
   3. at 4℃ overnight
4. Proceed with bacterial transformation

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