Isaguliants M G, Samoshin V V, Makeeva I V, Smirnova V D
Bioorg Khim. 1990 Jul;16(7):933-40.
A facile technique of manual oligonucleotide synthesis via H-phosphonate approach is developed. Syntheses carried out in pipette tips with siliconised glasswool filters take 3-3.5 min per cycle with 97-98% yields per condensation. The method was used to synthesize 12-55-mers: T7 and PL promoter regions, gene of the signal peptide of the E. coli OmpA protein, oligonucleotides coding for amino acid sequences 94-105 of preS1- and 133-143 of preS2-regions of hepatitis B virus, hybridisation probes, sequencing primers, oligonucleotides for site-directed mutagenesis, etc.
开发了一种通过H-膦酸酯法进行手动寡核苷酸合成的简便技术。在带有硅化玻璃棉过滤器的移液管尖端进行合成，每个循环耗时3 - 3.5分钟，每次缩合产率为97 - 98%。该方法用于合成12 - 55聚体：T7和PL启动子区域、大肠杆菌OmpA蛋白信号肽基因、编码乙肝病毒前S1区域94 - 105氨基酸序列和前S2区域133 - 143氨基酸序列的寡核苷酸、杂交探针、测序引物、用于定点诱变的寡核苷酸等。
