首页> 外国专利> Non-radioactive labelled single-stranded probe, method for manufacturing it and method for detecting a specified nucleotide sequence using this probe

Non-radioactive labelled single-stranded probe, method for manufacturing it and method for detecting a specified nucleotide sequence using this probe

机译:非放射性标记的单链探针,其制造方法和使用该探针检测特定核苷酸序列的方法

摘要

The invention relates to a new probe for detecting a specified nucleic acid sequence. It consists of a recombinant DNA containing an insert complementary to the nucleic acid sequence to be detected and nuclecic acid sequences derived from the DNA (+) of phage M13, the bases which participate in the composition of this recombinant DNA, in particular the thymine groups, being replaced by 5-bromouracil groups. The invention also relates to a method for manufacturing this probe, this method comprising the culturing of cell hosts, more especially E. coli, which are auxotrophic for thymine in a medium containing a limiting concentration of thymine and containing 5-bromouracil. After the requisite incubation time, the supernatant is removed and the probe is recovered from the bacterial cells.
机译:本发明涉及用于检测特定核酸序列的新探针。它由含有与待检测核酸序列互补的插入片段的重组DNA和源自噬菌体M13的DNA(+)的核酸序列组成,这些碱基参与了该重组DNA的组成,特别是胸腺嘧啶基团,被5-溴尿嘧啶组取代。本发明还涉及用于制造该探针的方法,该方法包括在含有有限浓度的胸腺嘧啶和含有5-溴尿嘧啶的培养基中培养对胸腺嘧啶营养缺陷的细胞宿主,尤其是大肠杆菌。经过必要的温育时间后,除去上清液,并从细菌细胞中回收探针。

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