"
Team:Newcastle/Ligation
From 2010.igem.org
(Difference between revisions)
Shethharsh08 (Talk | contribs) |
Shethharsh08 (Talk | contribs) (→Procedures) |
||
| Line 21: | Line 21: | ||
|3 | |3 | ||
|5 | |5 | ||
| - | | | + | |N/A |
|- | |- | ||
|'''10X BUFFER''' | |'''10X BUFFER''' | ||
Revision as of 08:48, 30 July 2010
| |||||||||||||
| |||||||||||||
Ligation
Procedures
Set up the ligation mix according:
| Reagents | 1:3(μl) | 1:5(μl) | Vector(μl) |
|---|---|---|---|
| Vector | 1 | 1 | 1 |
| Insert | 3 | 5 | N/A |
| 10X BUFFER | 1 | 1 | 1 |
| T4 Ligase | 1 | 1 | 1 |
| H2O | 4 | 2 | 7 |
| Total Volume | 10.0 | 10.0 | 10.0 |
For sticky end ligations incubate on the bench of 3-4 hrs or overnight
Notes
- For standard ligation 100-200ng of vector DNA is required - can be checked using nanodrop
- A ligation calcultion can be used however a 1:3 and a 1:5 ratio tends to be used instead.
   
|
    
|
