Purification of Small DNA Fragments

Procedure

1. Add 150 μL of TE, pH 8.0 to a 50 μL amplification reaction containing your PCR product.

2. Add 100 μL of 30% PEG 8000/ 30 mM MgCl2.

3. Vortex to mix thoroughly and centrifuge immediately at 10,000 G for 15 minutes at room temperature.

4. Carefully remove the supernatant.

5. Dissolve the pellet in 50 μL of TE, pH 8.0.

6. Check the quality and quantity of the recovered PCR product on an agarose gel.

Notes and Cautions

• Increasing the centrifugation time and/or speed will increase the yield.

• The pellet will be clear and nearly invisible.

• If this is for attB recombination, the concentration should remain greater than 10 ng/μL.