关于化学合成非放射性生物素化核酸杂交探针开发的研究。
Studies towards the development of chemically synthesized non-radioactive biotinylated nucleic acid hybridization probes.
作者信息
Al-Hakim A H, Hull R
出版信息
Nucleic Acids Res. 1986 Dec 22;14(24):9965-76. doi: 10.1093/nar/14.24.9965.
Abstract
Non-radioactive nucleic acid hybridization probes have been constructed in which the reporter group is long chain biotin chemically linked to a basic macromolecule (histone H1, cytochrome C or polyethyleneimine). The modified basic macromolecule which carries many biotin residues can, in turn, be covalently linked to nucleic acids (DNA) via the bifunctional cross-linking reagents, glutaraldehyde, 1,2,7,8-diepoxyoctane, bis (succinimidyl) suberate or bis (sulfonosuccinimidyl) suberate. This provides a very sensitive probe by which as little as between 10-50fg of target DNA can be visualized using dot-blot hybridization procedures in conjunction with avidin or streptavidin enzyme conjugates.
摘要
已构建了非放射性核酸杂交探针，其中报告基团是化学连接到碱性大分子（组蛋白H1、细胞色素C或聚乙烯亚胺）的长链生物素。携带许多生物素残基的修饰碱性大分子可通过双功能交联剂戊二醛、1,2,7,8 - 二环氧辛烷、辛二酸双（琥珀酰亚胺酯）或辛二酸双（磺基琥珀酰亚胺酯）与核酸（DNA）共价连接。这提供了一种非常灵敏的探针，通过斑点杂交程序结合抗生物素蛋白或链霉抗生物素蛋白酶偶联物，可检测低至10 - 50fg的靶DNA。
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