New reversed-phase high-performance liquid chromatographic column for oligonucleotide separation

✍ Scribed by Hiroyuki Moriyama; Yoshio Kato

• Publisher: Elsevier Science
• Year: 1988
• Tongue: English
• Weight: 434 KB
• Volume: 445
• Category: Article
• ISSN: 1873-3778
• DOI: 10.1016/s0021-9673(01)84525-8

✦ Synopsis

Modern reversed-phase chromatography (RPC) has been widely applied to the separation of nucleobases, nucleosides and nucleotides. Long-chain oligo-and polynucleotides, which are valuable tools in the genetic engineering process, have become easily available because of the rapid developmental of automated DNA synthesizers. The synthesized mixture commonly contains many compounds such as short chains so that oligo-and polynucleotides have to be rapidly and successfully purified from by-products after the synthetic process. It is important to obtain high quality oligonucleotides for gene manipulation.

High-performance liquid chromatography (HPLC) is extensively utilized for separating nucleic acids. For instance, gel filtration chromatography (GFC) is suitable for isolation of large molecules such as DNA fragmentsie3 and ion-exchange chromatography (IEC) is advantageous for separation of a wide range of nucleic acids4-6. On the other hand, RPC is also an useful method for separation of nucleic acid samples, mainly small compounds7-9.

Recently, a new reversed-phase chromatographic column, TSKgel OligoDNA RP (Tosoh, Tokyo, Japan) has become commercially available. This new column, which has a wide pore size and octadecyl groups bonded to silica gel, is designed for separation of medium-molecular-weight compounds such as oligo-and polynucleotides. We have evaluated it in terms of oligonucleotide separation by comparison with a conventional ODS column and the results are now described.