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PCR Purification

Materials

• 1.5 ml microcentrifuge tubes
• 2 ml collection tube
• QIAquick columns
• Qiagen Buffer PB
• Qiagen Buffer EB
• Qiagen Buffer PE
• DNA mixture from PCR

Protocol

1. Add 5 volumes of Qiagen Buffer PB to 1 volume of PCR product.
2. Put a QIAquick spin column into a 2ml collection tube.
3. Apply the mixture to the QIAquick column to bind the DNA and centrifuge for 30 to 60 seconds at 13,000 rpm.
4. Discard flow-through. Add 0.75 ml of Buffer PE to the spin column to wash and centrifuge for 30 to 60 seconds at 13,000 rpm.
5. Discard flow-through and centrifuge for another 1 minute at 13,000 rpm.
6. Place QIAquick spin column into a clean 1.5 ml microcentrifuge tube.
7. Add 50 µl Buffer EB to the center of the membrane of spin column and centrifuge for 1 minute at 13,000 rpm.
8. If analysing purified DNA using gel, add 1 volume of Loading Dye to 5 volumes of DNA.
9. Store the PCR product either at 4°C or at -20°C.

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