化学法合成生物素标记寡核苷酸探针

Chemical Synthesis of th Biotinylated Oligonucleotide Probes

生物素可通过氨烷基磷酰胺基团连接到寡核苷酸5'末端,反应在水溶液中进行,核苷酸的侧链基团不用保护。我们以这种化学法合成了生物素标记的寡核苷酸探针,其显色灵敏度达50pg,杂交灵敏度达0.4fmol,并与酶标生物素寡核苷酸探针进行了比较,也对两种不同显色体系作了比较。

Oligonucleotides were labeled with biotin at their 5' end through an am inoalkylphosphoramide arm group. The reaction was performed in aqueous solution and the side chains on nucleotides were not necessary to protect. The chemically synthesized biotin-labeled probes had a colorimetric sensitivity of 50 pg and a hybridization-one of 0.4 fmol. They could be prepared in large amount and were comparable to the enzymatically biotinlabeled probes after purification.