Qiagen QIAquick PCR Purification Kit Protocol
Add 5 volumes of Buffer PB to 1 volume of the PCR sample and mix.
It is not necessary to remove mineral oil or kerosene.
If pH indicator I has beein added to Buffer PB, check that the color of the mixture is yellow.
If the color of the mixture is orange or violet, add 10 µl of 3 M sodium acetate, pH 5.0, and mix.
The color of the mixture will turn to yellow.
Place a QIAquick spin column in a provided 2 ml collection tube.
To bind DNA, apply the sample to the QIAquick column and centrifuge for 30–60 s. Discard flow-through.
Place the QIAquick column back into the same tube.
To wash, add 0.75 ml Buffer PE to the QIAquick column and centrifuge for 30–60 s. 
Discard flow-through and place the QIAquick column back in the same tube.
Centrifuge the column for an additional 1 min.
IMPORTANT: Residual ethanol from Buffer PE will not be completely removed unless the flow-through is discarded before this additional centrifugation.
Place QIAquick column in a clean 1.5 ml microcentrifuge tube.
To elute DNA, add 50 µl Buffer EB (10 mM Tris·Cl, pH 8.5) or water (pH 7.0–8.5) to the center of the QIAquick membrane and centrifuge the column for 1 min.
Alternatively, for increased DNA concentration, add 30 µl elution buffer to the center of the QIAquick membrane, let the column stand for 1 min, and then centrifuge.
IMPORTANT: Ensure that the elution buffer is dispensed directly onto the QIAquick membrane for complete elution of bound DNA.
The average eluate volume is 48 µl from 50 µl elution buffer volume, and 28 µl from 30 µl elution buffer.
Elution efficiency is dependent on pH.
The maximum elution efficiency is achieved between pH 7.0 and 8.5.
When using water, make sure that the pH value is within this range, and store DNA at –20°C as DNA may degrade in the absence of a buffering agent.
The purified DNA can also be eluted in TE buffer (10 mM Tris·Cl, 1 mM EDTA, pH 8.0), but the EDTA may inhibit subsequent enzymatic reactions.
If the purified DNA is to be analyzed on a gel, add 1 volume of Loading Dye to 5 volumes of purified DNA.
Mix the solution by pipetting up and down before loading the gel.
Loading dye contains 3 marker dyes (bromophenol blue, xylene cyanol, and orange G) that facilitate estimation of DNA migration distance and optimization of agarose gel run time.
Refer to Table 2 (page 15) to identify the dyes according to migration distance and agarose gel percentage and type.
