Team:Kyoto/BindingAssays

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===Ligation===
===Ligation===
#Make 2µL of Mixture (the vector and the insert at 1 : 5-10) and Control (only the vector).  
#Make 2µL of Mixture (the vector and the insert at 1 : 5-10) and Control (only the vector).  

Revision as of 21:46, 4 October 2011

Ligation

  1. Make 2µL of Mixture (the vector and the insert at 1 : 5-10) and Control (only the vector).
  2. Add 5µL Ligation High, 1µL T4 Kinase, and 7µL MilliQ to create a solution.
  3. Incubate at 16℃ for 30 min. If the colonies of E.coli transformed with the Control

Restrictive Digestion

  1. Use EcoRI, XbaI, SpeI, PstI, (NEB)
  2. Mix the following.
    Sample 5µL
    10xBuffer 1µL
    Restriction Enzyme 0.1µL
    MilliQ -3.9µL
  3. Let stand for 2h at 37℃
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