Photocleavable biotin phosphoramidite for 5'-end-labeling, affinity purification and phosphorylation of synthetic oligonucleotides
Photocleavable biotin phosphoramidite for 5'-end-labeling, affinity purification and phosphorylation of synthetic oligonucleotides
复制标题
DOI：
10.1093/nar/24.2.361
复制
发表时间：
1996-01-15
影响因子：
14.9
通讯作者：
Rothschild, KJ
中科院分区：
文献类型：
--
作者：
Olejnik, J;KrzymanskaOlejnik, E;Rothschild, KJ
We report the design, synthesis and evaluation of a non-nucleosidic photocleavable biotin phosphoramidite (PCB-phosphoramidite) which provides a simple method for purification and phosphorylation of oligonucleotides. This reagent introduces a photocleavable biotin label (PCB) on the 5'-terminal phosphate of synthetic oligonucleotides and is fully compatible with automated solid support synthesis, HPLC analysis shows that the PCB moiety is introduced predominantly on full-length sequences and is retained during cleavage of the synthetic oligonucleotide from the solid support and during subsequent deprotection with ammonia. The full-length 5'-PCB-labeled oligonucleotide can then be selectively isolated from the crude oligonucleotide mixture by incubation with immobilized streptavidin, Upon irradiation with 300-350 nm light the 5'-PCB moiety is cleaved with high efficiency in
