"
Team:Kyoto/BindingAssays
From 2011.igem.org
(Difference between revisions)
| Line 3: | Line 3: | ||
{{Kyoto_WikiDesign}} | {{Kyoto_WikiDesign}} | ||
| + | =='''Binding Assays'''== | ||
===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 21:48, 4 October 2011
Binding Assays
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℃
