Septak M
PerSeptive Biosystems, Framingham, MA 01701, USA.
Nucleic Acids Res. 1996 Aug 1;24(15):3053-8. doi: 10.1093/nar/24.15.3053.
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 >> 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-[17mer] 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.
使用完全保护的固定在CPG上的单体、二聚体和三聚体寡核苷酸来研究寡核苷酸化学合成过程中的脱嘌呤作用。通过反相高效液相色谱分析监测相对于不受脱嘌呤影响的内部胸苷标准品，在酸暴露时间内寡核苷酸的消失情况。发现在二氯甲烷中，二氯乙酸（DCA）和三氯乙酸（TCA）的脱嘌呤半衰期为3%DCA >> 15%DCA > 3%TCA。为了理解DNA合成过程中脱嘌呤的影响，还测量了模型化合物DMT-dG-pT二聚体和DMT-[17聚体]混合碱基序列的去三苯甲基化动力学。这些结果提高了我们在寡核苷酸合成中适当平衡获得最大去三苯甲基化与最小脱嘌呤这两个相互矛盾目标的能力。
