Editing PEG8000-PCR-Purification

Purify PCR product using PEG 8000 

This protocol removes DNA less than 300 bp in size. 

1. Add 75 ÎŒl of 1X TE, pH 8.0 to a 25 ÎŒl PCR amplification reaction containing your PCR product. 

2. Add 50 ÎŒl of 30% PEG 8000/30 mM MgCl<sub>2</sub>. Vortex to mix thoroughly and centrifuge immediately at 10,000 x g for 15 minutes at room temperature. 

In most cases, centrifugation at 10,000 x g for 15 minutes results in efficient recovery of PCR products. To increase the amount of PCR product recovered, the centrifugation time may be extended or the speed of centrifugation increased. 

3. Carefully remove the supernatant. The pellet will be clear and nearly invisible. 

4. Dissolve the pellet in 25 ÎŒl of 1X TE, pH 8.0 (to concentration &gt;10 ng/ÎŒl). 

5. Check the quality and quantity of the recovered PCR product by running an aliquot on an agarose gel. 

[[Category:Protocols]]