Theisen P, McCollum C, Andrus A
Applied Biosystems Inc., Foster City, CA 94404.
Nucleic Acids Symp Ser. 1992(27):99-100. doi: 10.1002/chin.199302246.
A series of fluorescein phosphoramidites (FAM) have been synthesized for use on automated DNA synthesizers. After coupling of the FAM reagents to the 5' hydroxyl of the oligonucleotide on the DNA synthesizer, the excess reagent is removed by washing the solid support. The dye, and its linkage to the oligonucleotide, are stable during the conditions of DNA synthesis and cleavage/deprotection conditions. Purification is attained with the OPC (Oligonucleotide Purification Cartridge), a polystyrene based affinity matrix, which selectively retains hydrophobic oligonucleotide conjugates. Analysis by MicroGel capillary electrophoresis effectively separates fluorescent dye labelled oligonucleotides from unlabelled products.
已合成了一系列用于自动DNA合成仪的荧光素亚磷酰胺（FAM）。在DNA合成仪上，将FAM试剂与寡核苷酸的5'羟基偶联后，通过洗涤固相载体除去过量试剂。在DNA合成及切割/脱保护条件下，染料及其与寡核苷酸的连接是稳定的。使用基于聚苯乙烯的亲和基质OPC（寡核苷酸纯化柱）进行纯化，该基质可选择性保留疏水性寡核苷酸缀合物。通过微凝胶毛细管电泳分析可有效分离荧光染料标记的寡核苷酸与未标记产物。
