Daniel:Protocols/CentriSep

From ZhangLabWiki
Jump to navigation Jump to search

Centri-Sep Protocol[edit]

Back to Main

Princeton Separations Info: Centri Sep Protocol

Catalog No.

  • CS-900 (32 pack) $109
  • CS-901 (100 pack) $287
Use this protocol for Centri-Sep column purifications (Princeton Separations).  This protocol uses size exclusion to separate DNA components
from fluorophores post dye coupling.

Protocol[edit]

1. Hydrate in 800 uL ddH20; Allow at least 30 minutes for hydration, be sure to hydrate all the gel
2. Remove air bubbles by inverting and sharply tapping the column; stand up column and allow to settle
3. Drain the column into a wash tube (200-250 uL will drain)
4. Centrifuge at 750xg for 2 minutes (~3000 rpm; removes 300 uL fluid)
5. Transfer 20 uL reaction mixture to the top of the gel; do not disturb gel surface
6. Place column into collection tube and centrifuge at 750xg for 2 minutes
7. Dry sample in vacuum centrifuge