Dizdaroglu M, Simic M G, Schott H
J Chromatogr. 1980 Jan 25;188(1):273-9. doi: 10.1016/s0021-9673(00)88442-3.
Chemical synthesis of DNA fragments with a defined sequence is extremely long and complex. A simpler alternative is isolation of fragments from partially hydrolysed DNA. Mixtures of sequence isomers of pyrimidine oligonucleotides were recently isolated on a preparative scale from hydrolysates of depurinated herring sperm DNA. These, however, could not be further separated by conventional techniques. Using high-performance liquid chromatography (HPLC), sequence isomers of pyrimidine di-, tri- and tetranucleotides were successfully separated and sequenced. In the present work, separation and sequencing of sequence isomers of pyrimidine pentanucleotides by HPLC was achieved, demonstrating the applicability of this particular approach in preparation of desired DNA fragments.
化学合成具有特定序列的DNA片段极其漫长且复杂。一种更简单的替代方法是从部分水解的DNA中分离片段。最近，在制备规模上从脱嘌呤鲱鱼精子DNA的水解产物中分离出了嘧啶寡核苷酸的序列异构体混合物。然而，这些异构体无法通过传统技术进一步分离。使用高效液相色谱（HPLC），成功分离并测序了嘧啶二核苷酸、三核苷酸和四核苷酸的序列异构体。在本研究中，通过HPLC实现了嘧啶五核苷酸序列异构体的分离和测序，证明了这种特定方法在制备所需DNA片段中的适用性。
