Nucleic acid separation using immobilized metal affinity chromatography

A method for separating compounds comprising the steps of:

• passing a mixture of compounds including target polynucleotides comprising at least four non-shielded purine moieties, at least four non-shielded pyrimidine moieties, or mixtures thereof, through a column comprising an IMAC ligand, wherein the ligand is capable of differentially binding the polynucleotides;
• and
• eluting and collecting purified samples of the target polynucleotides substantially free from both protein contaminants and histidine tags.

A method for separating compounds comprising the step of:

• contacting a mixture comprising cell lysate or enzyme and a target polynucleotide including a non-shielded purine or pyrimidine moiety and a compound including a shielded purine or pyrimidine moiety with a solid composition including immobilized metal ions capable of binding compounds containing a non-shielded purine or pyrimidine moiety to form a supernatant liquid having a reduced amount of compounds including a non-shielded purine or pyrimidine moiety;
• wherein the compound including a non-shielded purine or pyrimidine moiety comprises a single stranded nucleic acid oligomer, or a single stranded nucleic acid polymer and the compounds including a shielded purine or pyrimidine moiety comprise double stranded nucleic acid oligomers or double stranded nucleic acid polymers;
• wherein the supernatant liquid contains less than or equal to 5% by weight compounds comprising a non-shielded purine or pyrimidine moiety and the solid composition is substantially free from both protein contaminants and histidine tags.

A method for separating compounds comprising the step of:

• contacting a mixture comprising cell lysate or enzyme comprising double-stranded DNA and additionally comprising RNA and/or DNA, said RNA and/or DNA containing single-stranded portions having a non-shielded purine or pyrimidine moiety, with a solid composition comprising immobilized metal ions capable of binding compounds having a non-shielded purine or pyrimidine moiety, to form a supernatant liquid having a reduced amount of RNA and/or DNA having single-stranded portions and a solid composition substantially free from both protein contaminants and histidine tags.

A method for separating compounds comprising the steps of:

• passing a solution comprising at least one polynucleotide, the polynucleotide containing single-stranded portions having at least four non-shielded purine or pyrimidine moieties through a column including an IMAC ligand,
• where the ligand is capable of differentially binding the polynucleotide without the presence of histidine tags;
• and collecting purified samples of each polynucleotide compound substantially free from both protein contaminants and histidine tags.