Kinetic studies on depurination and detritylation of CPG-bound intermediates during oligonucleotide synthesis
Kinetic studies on depurination and detritylation of CPG-bound intermediates during oligonucleotide synthesis
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DOI：
10.1093/nar/24.15.3053
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发表时间：
1996-08-01
影响因子：
14.9
通讯作者：
Septak, M
中科院分区：
文献类型：
--
作者：
Septak, M
完全保护的CPG固定化的单体，二聚体和三聚体寡核苷酸用于研究在化学合成寡核苷酸过程中的脱嘌呤。通过反相HPLC分析监测在酸暴露时间期间相对于未进行脱嘌呤的胸苷内标物的寡核苷酸的消失。二氯乙酸（DCA）和三氯乙酸（TCA）在二氯甲烷中的脱尿半衰期为3%DCA，远大于15%DCA> 3%TCA。为了理解脱嘌呤在DNA合成过程中的意义，还测量了模型化合物DMT-dG-pT二聚体和DMT-[17 mer]混合碱基序列的脱三苯甲基动力学。这些结果提高了我们在寡核苷酸合成中获得最大脱三苯甲基与最小脱嘌呤的矛盾目标的适当平衡的能力。
Fully protected CPG-immobilized monomer, dimer and trimer oligonucleotides were used to study depurination during the chemical synthesis of oligonucleotides. Disappearance of the oligonucleotide during acid exposure time relative to an internal thymidine standard not subject to depurination was monitored by reverse phase HPLC analysis. Depurination half-times obtained for dichloroacetic acid (DCA) and trichloroacetic acid (TCA) in methylene chloride were found to be 3% DCA much greater than 15% DCA > 3% TCA. In order to understand the implications of depurination during DNA synthesis, the detritylation kinetics of model compounds DMT-dG-pT dimer and DMT-[17 mer] mixed-base sequence were also measured. These results improve our ability to properly balance the contradictory goals of obtaining maximum detritylation with minimum depurination in oligonucleotide synthesis.