From 2011.igem.org
(Difference between revisions)
|
|
| Line 2: |
Line 2: |
| | {{Kyoto_Background}} | | {{Kyoto_Background}} |
| | {{Kyoto_WikiDesign}} | | {{Kyoto_WikiDesign}} |
| - |
| |
| - | =='''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===
| |
| - | <ol>
| |
| - | <li>Use EcoRI, XbaI, SpeI, PstI, (NEB)</li>
| |
| - | <li>Mix the following.
| |
| - | {|
| |
| - | |-
| |
| - | | Sample
| |
| - | | 5µL
| |
| - | |-
| |
| - | | 10xBuffer
| |
| - | | 1µL
| |
| - | |-
| |
| - | | Restriction Enzyme
| |
| - | | 0.1µL
| |
| - | |-
| |
| - | | MilliQ
| |
| - | | -3.9µL
| |
| - | |}
| |
| - | </li>
| |
| - | <li>Let stand for 2h at 37℃</li>
| |
| - | </ol>
| |
Latest revision as of 03:32, 5 October 2011
"
