"
Team:Kyoto/BindingAssays
From 2011.igem.org
(Difference between revisions)
Redsalmon (Talk | contribs)
(Created page with "===Ligation=== #Make 2µL of Mixture (the vector and the insert at 1 : 5-10) and Control (only the vector). #Add 5µL Ligation High, 1µL T4 Kinase, and 7µL MilliQ to create a ...")
Newer edit →
(Created page with "===Ligation=== #Make 2µL of Mixture (the vector and the insert at 1 : 5-10) and Control (only the vector). #Add 5µL Ligation High, 1µL T4 Kinase, and 7µL MilliQ to create a ...")
Newer edit →
Revision as of 21:43, 4 October 2011
Ligation
- Make 2µL of Mixture (the vector and the insert at 1 : 5-10) and Control (only the vector).
- Add 5µL Ligation High, 1µL T4 Kinase, and 7µL MilliQ to create a solution.
- Incubate at 16℃ for 30 min. If the colonies of E.coli transformed with the Control
Restrictive Digestion
- Use EcoRI, XbaI, SpeI, PstI, (NEB)
- Mix the following.
Sample 5µL 10xBuffer 1µL Restriction Enzyme 0.1µL MilliQ -3.9µL - Let stand for 2h at 37℃
