Promega 1-2
Wizard Genomic DNA Clean-Up System
1. Use one Wizard Mini-column for each sample.
2. Remove and set aside the plunger from a 3mL disposable syringe.
3. Attach one Syringe Barrel to the Luer-Lok extension of one Mini-column
4. Place this Mini-column into a 1.5mL micro-centrifuge tube.
5. Thoroughly mix the Wizard DNA Clean-Up Resin before removing an aliquot.
a. If crystals or aggregates are present, dissolve by warming the resin to 37oC
for 10 minutes.
b. The resin itself is insoluble.
c. Cool to 25-30oC before use.
6. Add 1mL of Wizard DNA Clean-Up Resin to a 1.5mL micro-centrifuge tube.
7. Add the sample to the Clean-Up Resin
a. The sample volume must be between 50 and 500ul.
b. If the sample volume is less than 50ul, bring the volume up to at least 50ul
with sterile water.
8. Mix by inversion.
9. Transfer the Wizard DNA Clean-Up Resin containing the bound DNA into the
Syringe Barrel attached to a Mini-column.
10. Insert the syringe plunger slowly
11. Gently push the slurry into the Mini-column with the syringe plunger.
12. Detach the syringe from the Mini-column
13. Remove the plunger from the syringe.
14. Reattach the Syringe Barrel to the Mini-column.
15. Add 2mL of 80% isopropanol into the syringe.
16. Reinsert the plunger
17. Gently push the solution through the Mini-column.
18. Remove the Syringe Barrel
19. Transfer the Mini-column to a new sterile 1.5mL micro-centrifuge tube.
20. Centrifuge the Mini-column at maximum speed in a micro-centrifuge for 2
minutes to dry the resin.
Promega 2-2
21. Transfer the Mini-column to a new 1.5mL micro-centrifuge tube.
22. Apply 50ul of pre-warmed (65-70oC) water to the Mini-column
23. Wait 1 minute.
24. Centrifuge the Mini-column for 20 seconds at maximum speed in a microcentrifuge to elute the bound DNA.
25. Remove and discard the Mini-column.
26. Store the purified DNA at –20oC