DNA微阵列原位合成中的氧化剂体系改进  

Improvement of Oxidizing Agent for DNA Microarray in situ Synthesis

作　　者：聂立波[1] 肖鹏峰[1] 汤建新[1] 谭美君[2] 陈洪[2] 何农跃[1] 

机构地区：[1]东南大学分子与生物分子电子学教育部重点实验室,南京210096 [2]株洲工学院包装印刷学院,株洲412008

出　　处：《高等学校化学学报》2004年第9期1676-1678,共3页Chemical Journal of Chinese Universities

基　　金：国家自然科学基金 (批准号 :60 0 710 0 1);国家高技术研究发展计划 (批准号 :2 0 0 3 AA3 0 2 2 70 );教育部跨世纪优秀人才培养计划资助

摘　　要：The phosphorous amide approach is of the highest coupling efficiency among the DNA in situ synthesis approaches at present. The strict anhydrous condition is required in the process of DNA in situ synthesis, and the DNA synthesis in the molecular stamp method must be performed in the anhydrous glove box. Because various volatile reagents remain in the glove box, the reagent gases invade into the coupling reaction system so that the coupling efficiency is decreased. This decrease is caused by the pyridine contained in the oxidizing agent. An improved oxidizing agent system of I 2/Ac 2O/AcOH/THF for the synthesis of oligodeoxy-nucleotides was reported. Using this oxidizing agent, 16-mer oligodeoxynucleotide probes were successfully synthesized on the CPG(controlled pore size glass) or the modified glass slide and a single step coupling efficiency of 98.2% was achieved. The synthesized DNA probes were hybridized with fluorescence and gold-labeled target oligodeoxynucleotides respectively. The fluorescence intensity and gray level for the synthesized probes in the I 2/Ac 2O/AcOH/THF oxidizing agent were similar to those in the traditional phosphorous amide approach possessing a water- and pyridine-containing oxidizing agent I 2/H 2O/Pyridine/THF. The reported oxidizing agent will make the contact printing approach to in situ synthesized DNA microarrays more feasible by eliminating the decrease of coupling efficiency which is because the cross reagent interfering arises from pyridine and water.The phosphorous amide approach is of the highest coupling efficiency among the DNA in situ synthesis approaches at present. The strict anhydrous condition is required in the process of DNA in situ synthesis, and the DNA synthesis in the molecular stamp method must be performed in the anhydrous glove box. Because various volatile reagents remain in the glove box, the reagent gases invade into the coupling reaction system so that the coupling efficiency is decreased. This decrease is caused by the pyridine contained in the oxidizing agent. An improved oxidizing agent system of I 2/Ac 2O/AcOH/THF for the synthesis of oligodeoxy-nucleotides was reported. Using this oxidizing agent, 16-mer oligodeoxynucleotide probes were successfully synthesized on the CPG(controlled pore size glass) or the modified glass slide and a single step coupling efficiency of 98.2% was achieved. The synthesized DNA probes were hybridized with fluorescence and gold-labeled target oligodeoxynucleotides respectively. The fluorescence intensity and gray level for the synthesized probes in the I 2/Ac 2O/AcOH/THF oxidizing agent were similar to those in the traditional phosphorous amide approach possessing a water- and pyridine-containing oxidizing agent I 2/H 2O/Pyridine/THF. The reported oxidizing agent will make the contact printing approach to in situ synthesized DNA microarrays more feasible by eliminating the decrease of coupling efficiency which is because the cross reagent interfering arises from pyridine and water.

关 键 词：DNA芯片 接触压印 试剂互串 氧化剂 偶联效率 

分 类 号：O657[理学—分析化学]