"
Team:Tuebingen/Notebook/Protocols/control-restriction
From 2013.igem.org
(Difference between revisions)
| Line 70: | Line 70: | ||
</div> | </div> | ||
| - | |||
| - | |||
| - | |||
| - | |||
| - | |||
| - | |||
| - | |||
| - | |||
| - | |||
| - | |||
| - | |||
<!-- FOOTER --- IMPORTANT FOR NAVMENU-LENGTH!!! --> | <!-- FOOTER --- IMPORTANT FOR NAVMENU-LENGTH!!! --> | ||
Revision as of 02:01, 4 October 2013
Control Restriction Digest
Back to Protocols
Reagents
| 1.5 µL | 10x NEBuffer 2 |
| 0.15 µL | 100x BSA |
| 3 µL | DNA from Mini-Prep (c > 50 ng/µL) |
| 0.15 µL | EcoRI or XbaI |
| 0.15 µL | PstI or SpeI |
| to 15 µL | Aqua dest. |
Procedure
- Around noon, mix all reagents in an 1.5 mL Eppendorf-tube and incubate at 37 °C until afternoon.
- Heat inactivate restriction enzymes at 80°C for 20 min.
- Add 1/10 volume of 10X Antarctic Phosphatase and mix. Incubate at 37 °C for 15 min for 5' extensions.
- Heat inactivate phosphatase at 70 °C for 5 min.
- Run control-gel in order to check a ligation / transformation.
