An immobilized metal affinity chromatography (IMAC) method for separating
and/or purifying compounds containing a non-shielded purine or pyrimidine
moiety or group such as nucleic acid, presumably through interaction with
the abundant aromatic nitrogen atoms in the purine or pyrimidine moiety.
The method can also be used to purify compounds containing purine or
pyrimidine moieties where the purine and pyrimidine moieties are shielded
from interaction with the column matrix from compounds containing a
non-shielded purine or pyrimidine moiety or group. Thus, double-stranded
plasmid and genomic DNA, which has no low binding affinity can be easily
separated from RNA and/or oligonucleotides which bind strongly to
metal-charged chelating matrices. IMAC columns clarify plasmid DNA from
bacterial alkaline lysates, purify a ribozyme, and remove primers and
other contaminants from PCR reactions. The metal ion affinity of yeast
RNA decreases in the order: copper (II), nickel (II), zinc (II), and
cobalt (II).