Seibl R, Höltke H J, Rüger R, Meindl A, Zachau H G, Rasshofer R, Roggendorf M, Wolf H, Arnold N, Wienberg J
Boehringer Mannheim GmbH, Biochemisches Forschungszentrum Penzberg, Tutzing.
Biol Chem Hoppe Seyler. 1990 Oct;371(10):939-51. doi: 10.1515/bchm3.1990.371.2.939.
The digoxigenin-based non-radioactive DNA labeling and detection system was applied in various hybridization protocols using digoxigenin-labeled probes obtained by enzymatic incorporation of Dig-[11]-dUTP. In genomic blots single-copy genes (human tissue-type plasminogen activator, constant part of immunoglobulin kappa light chain) can be detected with only 0.5 to 5 micrograms human DNA depending on the type of probe and the length of the hybridizing region. Due to its high sensitivity and specificity, the digoxigenin system is also appropriate for colony-, plaque-, and in situ hybridizations with metaphase chromosome spreads and fixed cells. Especially in the latter applications it is of great advantage, that with the digoxigenin system any significant background or unspecific side reactions with biological materials are avoided.
基于地高辛配体的非放射性DNA标记和检测系统被应用于各种杂交实验方案中，这些方案使用通过酶促掺入地高辛配体-[11]-dUTP获得的地高辛配体标记探针。在基因组印迹中，根据探针类型和杂交区域长度，仅用0.5至5微克人类DNA就可以检测到单拷贝基因（人类组织型纤溶酶原激活剂、免疫球蛋白κ轻链恒定区）。由于其高灵敏度和特异性，地高辛配体系统也适用于与中期染色体铺展和固定细胞的菌落、噬菌斑和原位杂交。特别是在后者的应用中，地高辛配体系统具有很大的优势，即避免了与生物材料发生任何明显的背景或非特异性副反应。
