The QIAprep PCR Purification Kit is intended to purify single or double stranded DNA fragments from PCR reactions. PCR fragments ranging in size from 100 bp to 10 kb can be purified from primers, enzymes, and salts. The kit uses silica-gel-membrane technology to bind DNA in a column while removing contaminants from the sample. After washing off the impurities, the DNA is eluted into a clean microcentrifuge tube.
The procedure is extremely quick (from start to finish takes only 10-15 minutes). Simply add 5 volumes of Buffer PB to 1 volume of PCR sample and mix; you don’t even need to remove the mineral oil. Pipette the solution into a QIAspin column and centrifuge for 1 minute. The DNA will bind to the column and the impurities will fall out in the flow-through. Next, wash the column with Buffer PE to enhance the purity of the DNA. Finally, elute DNA into a clean 1.5 mL microcentrifuge tube. Since maximum elution efficiency occurs at a pH of 7.0-8.5, if eluting with water rather than the supplied elution buffer, be sure that water has a pH within this range. Also, if eluting in water, store DNA at –20ºC, since without a buffer, the DNA may degrade at 4ºC.
We have had good success with this kit. The instructions are very easy to follow and all the necessary reagents are supplied, except ethanol. The only equipment required is a small tabletop centrifuge or vacuum manifold. I would recommend this kit to anyone who wants to clean up a PCR product in a quick, efficient manner.
Graduate Student
Department of Physiology
Emory University