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Chemically synthesized non-radioactive biotinylated long-chain nucleic acid hybridization probes

机译:化学合成的非放射性生物素化长链核酸杂交探针

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pA new method for the chemical labelling of nucleic acid with biotin to produce non-radioactive probes has been developed. NN9-Bis-(3-aminopropyl)butane-1,4-diamine (spermine) and long-chain diamino compounds (diaminohexane, diaminodecane and diaminododecane) were linked covalently to biotin and the resultant conjugates were attached to nucleic acid by using a cross-linking reagent (glutaraldehyde or diepoxyoctane). Iodoacetylation and biotinylation of the long-chain diamino compounds produced modified biotinylated conjugates that can be linked to DNA without the use of a cross-linking reagent. These types of probes attach one biotin molecule to each linker arm of spermine, diamino and iodoacetylated amino derivatives. Such probes have long linker arms separating the biotin moiety from the hybridization sites of the nucleic acid. These probes can detect 10 pg of target DNA by dot-blot hybridization./p
机译:已经开发了一种用生物素化学标记核酸以生产非放射性探针的新方法。 NN9-双-(3-氨基丙基)丁烷-1,4-二胺(精胺)和长链二氨基化合物(二氨基己烷,二氨基癸烷和二氨基十二烷)共价连接至生物素,并通过使用交叉将所得的缀合物连接至核酸-连接试剂(戊二醛或二环氧辛烷)。长链二氨基化合物的碘乙酰化和生物素化产生了修饰的生物素化缀合物,该缀合物可以与DNA连接而无需使用交联剂。这些类型的探针将一个生物素分子连接到精胺,二氨基和碘乙酰化氨基衍生物的每个连接臂上。这样的探针具有长的连接臂,其将生物素部分与核酸的杂交位点分开。这些探针可通过斑点印迹杂交检测10 pg的目标DNA。

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