RP-labeled oligonucleotide ion pair chromatography_0.doc

 PAGE \* MERGEFORMAT 18 RP-labeled oligonucleotide ion pair chromatography Study: High-Yun Li Peng Jing Hua Wu Liqing Shengling Hui Fu Boqiang [Abstract] labeled oligonucleotide established reversed-phase ion pair chromatography method to optimize the mobile phase concentration of acetic acid and triethylamine (0 ~ 0.15 mol / L, pH 4.5 ~ 7.0 and the elution strength of chromatographic conditions. On the 5 mer , 10 mer and 15 mer non-labeled and 5 ‘carboxy fluorescein (5’FAM labeled oligonucleotide comparative analysis of the reservation to study the fluorescent oligonucleotide retention mechanism and separation of other commonly used fluorescent probes TaqManTM labeled oligonucleotides. The results show that fluorescently labeled oligonucleotides of different lengths in the 0.01 mol / L triethylamine acetate, pH 7.0 under the conditions of maximum separation. fluorescently labeled oligonucleotide and unlabeled oligonucleotide reserved acid were significantly different between the two can be separated completely. In the context of a certain length of non-labeled oligonucleotides increased with length of retention time of growth, on the contrary, the length of fluorescently labeled oligonucleotides increase the retention time decreased. fluorescent dye hydrophobicity of its labeled oligonucleotide retention in reversed phase column in a greater impact, the stronger the hydrophobic fluorescent dye, the labeled oligonucleotides longer retention time. But the degree of influence of hydrophobic oligonucleotide labeled with nucleotide length increased gradually smaller. [Keywords:] fluorescently labeled oligonucleotides, fluorescent dyes, reversed-phase ion pair chromatography, retention 1 Introduction The completion of the Human Genome Project to promote the nucleic acid-based clinical diagnosis, forensic DNA identification, inspection and quarantine technology, so the qualitative and quantitative analysis of nucleic acids is becoming increasingly im