Team:Kyoto/BindingAssays

From 2011.igem.org

Binding Assays

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℃