Schmitz G G, Walter T, Seibl R, Kessler C
Boehringer-Mannheim GmbH, Biochemical Research Center, Penzberg, Federal Republic of Germany.
Anal Biochem. 1991 Jan;192(1):222-31. doi: 10.1016/0003-2697(91)90212-c.
A procedure for the nonradioactive labeling of oligonucleotides with the hapten digoxigenin (DIG) has been developed. The label is introduced by enzymatic tailing of the 3'-end. Two different modified nucleotides were applied. DIG-dUTP allows the synthesis of hapten-modified oligonucleotides with longer tails containing several DIG molecules, whereas the novel compound DIG-ddUTP leads to the addition of only a single DIG hapten. The efficiency of the labeling reactions with respect to variation of the different parameters was analyzed and data on application of labeled oligonucleotide probes to different blot formats are shown.
已开发出一种用半抗原地高辛（DIG）对寡核苷酸进行非放射性标记的方法。通过3'端的酶促加尾引入标记。应用了两种不同的修饰核苷酸。DIG-dUTP可合成带有较长尾巴且包含几个DIG分子的半抗原修饰寡核苷酸，而新型化合物DIG-ddUTP只会添加单个DIG半抗原。分析了标记反应在不同参数变化方面的效率，并展示了标记寡核苷酸探针应用于不同印迹形式的数据。
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