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Detection probe for leptocylindrus danicus

A technology for detecting probes and Cylindrophyllum, applied in DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of large fluctuations in detection results, difficult operation, and small number of capture probes, so as to facilitate the search and implementation , good probe applicability and high stability

Inactive Publication Date: 2014-01-29
OCEAN UNIV OF CHINA
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  • Abstract
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Problems solved by technology

Its main disadvantages are: first, because the length of the capture probe is only 10-30 bp, the number of capture probes that can effectively distinguish close relatives of microalgae is very small; second, it is necessary to prevent RNA degradation during the entire detection process, However, various steps such as washing and antibody reaction can easily cause RNA degradation, so the operation is difficult and the test results fluctuate greatly

Method used

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  • Detection probe for leptocylindrus danicus
  • Detection probe for leptocylindrus danicus
  • Detection probe for leptocylindrus danicus

Examples

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Embodiment

[0030] Example: Qualitative and quantitative detection of Cylindrocystis danishensis by using S1 enzyme protection analysis for rRNA and sandwich hybridization technology.

[0031] In this example, the combination of S1 enzyme protection analysis technology and sandwich hybridization technology is used to realize the qualitative and quantitative detection of marine red tide organism Leptocylindrus danicus. The steps are as follows:

[0032] 1.1 The search for the specific region of Cylindrocystis danishensis rRNA and the design and synthesis of S1 enzyme protection analysis probe

[0033] By determining the 18S rRNA sequence of the ribosomal small subunit of Cylindrophyllum danishensis and performing multiple sequence comparisons with the rRNA sequences of other algae, it was found that the 30-100 region of the small subunit rRNA was different from other algae, and the sequence of this region was determined to be a characteristic sequence ; Design and synthesize the S1 enzyme ...

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Abstract

The invention discloses a detection probe for leptocylindrus danicus, which belongs to the field of nucleic acid detection. Specifically aiming at a rRNA particular area of leptocylindrus danicus, a set of probes taking a S1 enzyme protection analysis probe as a core is formed, and in combination with S1 enzyme protection analysis and sandwich hybridization technologies, the qualitative and quantitative detection on leptocylindrus danicus rRNA is realized. The detection probe comprises three correlated oligonucleotide probes, namely, a S1 enzyme protection analysis probe, a capture probe, and a signal probe or a connecting probe. The probe disclosed by the invention is good in applicability, and greatly facilitates the finding and implementation of specific probes. The stability of the detection probe disclosed by the invention is far higher than that in the sandwich hybridization technology.

Description

technical field [0001] The invention relates to a group of oligonucleotide probes for algae detection, in particular to specific probes for qualitative and quantitative detection of Cylindrical algae. It belongs to the field of nucleic acid detection. Background technique [0002] Leptocylindrus danicus is a common coastal species with a very wide distribution. It is distributed in the Arctic Ocean, European seas, the Mediterranean Sea, the Atlantic and Pacific coasts of North America, the coasts of Japan and India, and the Bohai Sea to the South my country Sea in China. This species is a good bait for marine secondary producers, and it is also a common red tide organism in my country's coastal waters and a good pollution indicator. In economical seaweed farming areas, it often competes with kelp and laver for nutrients. Cause economic algae to change color, even rot and lose edible and commercial value. Therefore, it is of great significance to the marine aquaculture indus...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6834
Inventor 米铁柱王国善谭俊哲
Owner OCEAN UNIV OF CHINA
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