Efremov D G, Dimovski A J, Efremov G D
Department of Hematology, Faculty of Medicine, Skopje, Yugoslavia.
Hemoglobin. 1991;15(6):525-33. doi: 10.3109/03630269109027900.
A simple procedure for nonradioactive labeling of oligonucleotides has recently been developed (1). It consists of 3' end labeling of oligonucleotides with terminal transferase by incorporation of a single digoxigenin labeled dideoxy uridine triphosphate. We used these oligonucleotides for allele specific oligomer hybridization of polymerase chain reaction amplified DNA, followed by an enzyme-linked immunoassay and subsequent enzyme-catalyzed color reaction. We compared this procedure with the standard radioactive oligonucleotide hybridization technique through the detection of the most common Mediterranean beta-thalassemia mutations. This procedure was also used for the confirmation of a new mutation at position -87 (C----A) (2) of the beta-globin gene and for the subsequent family analysis.
最近开发了一种用于寡核苷酸非放射性标记的简单方法(1)。它包括通过掺入单个地高辛配基标记的双脱氧尿苷三磷酸，用末端转移酶对寡核苷酸进行3'末端标记。我们将这些寡核苷酸用于聚合酶链反应扩增DNA的等位基因特异性寡聚物杂交，随后进行酶联免疫测定和后续的酶催化显色反应。通过检测最常见的地中海β-地中海贫血突变，我们将该方法与标准放射性寡核苷酸杂交技术进行了比较。该方法还用于确认β-珠蛋白基因-87位（C→A）的一个新突变(2)，并进行后续的家系分析。
