Purification of PCR Products using 'Promega Wizard Preps'TM

[Promega® PCR Preps DNA Purification System, Promega UK Ltd, Delta House, Chilworth Research Centre, Southampton SO16 7NS, UK]

Note: It is advisable to prepare all tubes/columns and syringes before use.

1. Transfer PCR reaction mixture (after checking on the agarose gel) to a 1.5 ml tube (take care not to take up any mineral oil) .

2. Add 100 µl of 'Direct Purification Buffer' and mix.

3. Add 1 ml of 'Resin' and mix three times over one minute.

4. Prepare one 'Wizard Prep mini-column' for each PCR product.
a) Remove the plunger from a 2ml disposable syringe.
b) Attach syringe to mini-column (lock together).
c) Rest on the top of a 1.5 ml tube which has had it's lid removed.

5. a) Pipette the Resin/PCR mix into the syringe barrel (from step 3).
b) Insert the syringe plunger slowly and push the mix into the mini-column, letting liquid go to waste.

6. a) Remove syringe from mini-column, remove plunger, and re-attach syringe to the mini-column.
b) Pipette 2 ml of 80% isopropanol into the syringe.
c) Insert the syringe plunger and push the isopropanol through the mini-column to waste.

7. Remove the syringe and microfuge for 20 s at 12000 rpm to dry resin.

8. a) Transfer mini-column to a new 1.5 ml tube, and add 50 µl of DEPC water.
b) Leave for approximately one min (30 min maximum), and microfuge for 20 s at 12000 rpm.

9. Store at -20°C in a 0.75 ml tube.