IGEM:Harvard/2007/Protocols/Nucleotide Removal


 * 1) Add 10 volumes of Buffer PN to 1 volume of the reaction sample and mix.
 * 2) Place a QIAquick spin column in a provided 2 ml collection tube.
 * 3) To bind DNA, apply the sample to the QIAquick column and centrifuge for 1 min at 6000 rpm.
 * 4) Discard the flow-through and place QIAquick column back into the same tube.
 * 5) To wash QIAquick column, add 750 µl of Buffer PE and centrifuge for 1 min at 6000 rpm.
 * 6) Discard the flow-through and place the QIAquick column back in the same tube, which should be empty. Centrifuge for an additional 1 min at 13,000 rpm (~17,900 x g).
 * 7) Place the QIAquick column in a clean 1.5 ml microcentrifuge tube.
 * 8) To elute DNA, add 100–200 µl of Buffer EB (10 mM Tris·Cl, pH 8.5) or H2O to the center of the QIAquick membrane and centrifuge the column for 1 min at 13,000 rpm (~17,900 x g). Alternatively, for increased DNA concentration, add 30–50 µl elution buffer to the center of the QIAquick membrane, let the column stand for 1 min, and then centrifuge.