Gebeyehu G, Rao P Y, SooChan P, Simms D A, Klevan L
Nucleic Acids Res. 1987 Jun 11;15(11):4513-34. doi: 10.1093/nar/15.11.4513.
A series of dATP and dCTP nucleotide analogs have been synthesized which are modified by attachment of aliphatic linkers containing a functional group to the amino-nitrogen at the hydrogen bonding positions of the bases, that is, at the 6-position of adenine and the 4-position of cytosine. These nucleotides are incorporated into DNA probes by standard nick-translation protocols. DNA probes labeled with biotin derivatives of these nucleotides are effectively hybridized to target DNA sequences and can be detected by a streptavidin and calf intestinal alkaline phosphatase conjugate with a sensitivity (0.25 pg DNA) sufficient for reproducible and rapid detection of single copy genes in a Southern blot of mammalian DNA. Also, a procedure has been developed to allow reprobing of nylon filters that have been hybridized with biotinylated probes and developed with the streptavidin/alkaline phosphatase conjugate and a standard dye system.
已经合成了一系列dATP和dCTP核苷酸类似物，这些类似物通过在碱基的氢键位置（即腺嘌呤的6位和胞嘧啶的4位）将含有官能团的脂肪族连接体连接到氨基氮上而进行修饰。这些核苷酸通过标准的缺口平移方案掺入DNA探针中。用这些核苷酸的生物素衍生物标记的DNA探针能有效地与靶DNA序列杂交，并且可以通过链霉亲和素和小牛肠碱性磷酸酶缀合物进行检测，其灵敏度（0.25 pg DNA）足以在哺乳动物DNA的Southern印迹中对单拷贝基因进行可重复且快速的检测。此外，还开发了一种程序，用于对已与生物素化探针杂交并用链霉亲和素/碱性磷酸酶缀合物和标准染料系统显影的尼龙滤膜进行再次检测。
