Ruth J L
US Fish and Wildlife Forensics Laboratory, Ashland, OR 97520, USA.
Mol Biotechnol. 1996 Oct;6(2):163-78. doi: 10.1007/BF02740771.
The simple use of nonisotopic hybridization probes to detect complementary sequences provides valuable information in a large number of research and commercial applications. In hybridization assays, the four "S's (speed, simplicity, sensitivity, and specificity) are important criteria for determining the choice of probe and label. The direct chemical combination of synthetic oligonucleotide probes and enzyme labels offer advantages unmatched by other approaches, with the oligonucleotide providing rapid hybridization and high specificity, and the direct enzyme label providing simple and sensitive detection. Such oligonucleotide-enzyme conjugates ("oligozymes") can be used in a variety of hybridization and detection formats, including dot blots, Southern/northern blots, in situ, and solution hybridization/capture schemes. The practical synthesis and use of such oligozymes are summarized.
使用非同位素杂交探针检测互补序列,操作简单,在大量研究和商业应用中都能提供有价值的信息。在杂交分析中,四个“S”(速度、简便性、灵敏度和特异性)是决定探针和标记选择的重要标准。合成寡核苷酸探针与酶标记的直接化学结合具有其他方法无法比拟的优势,寡核苷酸能实现快速杂交和高特异性,直接酶标记则能提供简单灵敏的检测。这种寡核苷酸 - 酶缀合物(“寡酶”)可用于多种杂交和检测形式,包括斑点印迹、Southern/ Northern印迹、原位杂交以及溶液杂交/捕获方案。本文总结了此类寡酶的实际合成和应用方法。