A method for preparing long dna probes containing multiple repeating units

A technology of repeating units and long probes, applied in the field of oligonucleotide probe preparation, can solve the problems of limiting the magnification of bDNA structure and lower connection efficiency, and achieve the effect of increasing the magnification and increasing the sensitivity
CN108728430BActive Publication Date: 2022-04-05沈阳中科赛尔生物科技有限公司
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
沈阳中科赛尔生物科技有限公司
Publication Date
2022-04-05

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Abstract

The invention relates to the preparation of oligonucleotide probes, in particular to a method for preparing long DNA probes containing multiple repeating units. Using blunt-end endonuclease and nicking endonuclease to simultaneously cut the plasmid containing multiple repeat unit DNA double strands, and then use the principle of lambda exonuclease to specifically cut, after cutting, separate by electrophoresis to obtain multiple repeat units Single-stranded DNA of interest for the unit. The method of the present invention is suitable for preparing DNA probes of all sequence components, especially for sequences that are difficult to perform PCR amplification, such as those containing multiple repeating units, and the length of the prepared probe can reach 800nt, solving the problem of multiple repeating unit DNA Technical issues in the preparation of long probes.
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Description

technical field

[0001] The invention relates to the preparation of oligonucleotide probes, in particular to a method for preparing long DNA probes containing multiple repeating units. Background technique

[0002] Lambda exonuclease is a highly persistent exonuclease that selectively cleaves phosphorylated single strands from the 5'-3' end of double-stranded DNA. The most suitable substrate for Lambda exonuclease is 5' phosphorylated blunt-ended double-stranded DNA. It uses the 3'-end nucleic acid strand as a template to cut sequentially from the 5' end of the DNA, and forms at the center of both ends. single chain 【1】 . Another characteristic of Lambda exonuclease is that it cannot cut from the 5' end of the nick (nicking) site and the gap site 【2】 .

[0003] Branched-DNA signal amplification technology (branched-DNA, bDNA) is a nucleic acid hybridization signal amplification technology introduced by Chiron Company. This detection method has the characteristics of high ...

Claims

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