将硫醇特异性探针连接到合成寡脱氧核糖核苷酸3'末端的有效方法。
Efficient methods for attachment of thiol specific probes to the 3'-ends of synthetic oligodeoxyribonucleotides.
作者信息
Zuckermann R, Corey D, Schultz P
出版信息
Nucleic Acids Res. 1987 Jul 10;15(13):5305-21. doi: 10.1093/nar/15.13.5305.
Abstract
Methodology is described for the synthesis of DNA oligomers containing a free 3'-thiol group which can be selectively crosslinked with a wide variety of probes. This chemistry is compatible with both phosphotriester and phosphoramidite solid phase chemistry. Moreover, the sulphydryl group is introduced into the 3'-nucleoside solid support linkage prior to oligonucleotide synthesis. Consequently, no additional coupling steps are required after oligonucleotide synthesis, and isolation of the 3'-thiol oligonucleotide requires only one additional deprotection step. Cross-linking of the thiol-containing oligonucleotide to a fluorescent probe was carried out with high selectivity, in high yield, and under mild conditions.
摘要
本文描述了一种合成含有游离3'-硫醇基团的DNA寡聚物的方法，该基团可与多种探针进行选择性交联。这种化学方法与磷酸三酯和亚磷酰胺固相化学均兼容。此外，在寡核苷酸合成之前，巯基被引入到3'-核苷固相支持物连接中。因此，寡核苷酸合成后不需要额外的偶联步骤，分离3'-硫醇寡核苷酸仅需一个额外的脱保护步骤。含硫醇的寡核苷酸与荧光探针的交联反应具有高选择性、高产率且在温和条件下进行。
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