SYBR-Green fluorescent quantitative RT-PCR (reverse transcription-polymerase chain reaction) detection method for Microcystis aeruginosa phycophages in water environment

A microcystis green algae phagocytosis and a microcystis green algae phagocytosis technology are applied in the field of molecular biology and can solve the problem that there are not many methods for real-time quantitative changes of algae phages.

Inactive Publication Date: 2012-03-21
KUNMING UNIV OF SCI & TECH
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Problems solved by technology

But there are not many ways to study the real-time changes in the number of cyanophages

Method used

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  • SYBR-Green fluorescent quantitative RT-PCR (reverse transcription-polymerase chain reaction) detection method for Microcystis aeruginosa phycophages in water environment
  • SYBR-Green fluorescent quantitative RT-PCR (reverse transcription-polymerase chain reaction) detection method for Microcystis aeruginosa phycophages in water environment
  • SYBR-Green fluorescent quantitative RT-PCR (reverse transcription-polymerase chain reaction) detection method for Microcystis aeruginosa phycophages in water environment

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Embodiment 1

[0035] The experimental methods not indicating specific conditions in the following methods are usually according to conventional conditions such as the molecular cloning of Sambrook et al.: Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989) or the genetic engineering experimental technique of Peng Xiuling et al. (Science Technical Press), or as recommended by the manufacturer. The gene sequences of cyanophages were obtained from the Molecular Biology Database (NCBI).

[0036] 1. PCR primer design

[0037] According to the literature and combined with the BALST sequence alignment results in Genebank, refer to the tail sheath protein coding gene (g91) of Microcystis aeruginosa cyanophage Ma-LMM01, NCBI sequence number is AB242261, and use Primer Premier 5.0 and Oligo6 software to design primers . The quantitative PCR external reference standard primers SH-WF and SH-WR, quantitative primers SH-RTF and SH-RTF (Table 1) were designed, and the designed pr...

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Abstract

The invention discloses an SYBR-Green fluorescent quantitative RT-PCR (reverse transcription-polymerase chain reaction) detection method for Microcystis aeruginosa phycophages in a water environment, belonging to the field of molecular biology. In the invention, based on the g91 segment of the tail sheath protein gene of Microcystis aeruginosa phycophages Ma-LMM01, Premier5.0 and Oligo6 software are used to design quantitative PCR external standard primers SH-WF and SH-WR and quantitative PCR primers SH-RTF and SH-RTR, and the primers are used to detect the quantity of Microcystis aeruginosa phycophages in a water environment, thereby analyzing the law that the quantity of Microcystis aeruginosa phycophages in the water varies with time and space.

Description

technical field [0001] The invention relates to a method for quantitatively detecting and analyzing Microcystis aeruginosa cyanophages in a water body environment by using SYBR-Green fluorescence quantitative RT-PCR technology, belonging to the field of molecular biology. Background technique [0002] Cyanobacteria are a class of prokaryotic photoautotrophs with some characteristics of bacteria, so they are also called Cyanobacteria. Viruses that infect cyanobacteria are called cyanophages because they are very similar to bacteriophages. Cyanobacteria are the main cause of algal blooms. In recent years, with the development of economy, the increase of human activities and the increase of eutrophication in water bodies, the occurrence of cyanobacteria "bloom" has become increasingly serious. Cycophages exist in large numbers in both ocean and freshwater, and their content changes significantly before and after cyanobacteria blooms subside. As a potential specific biological...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68G01N21/64
Inventor 刘丽刘腾腾夏雪山向安冯悦魏大巧
Owner KUNMING UNIV OF SCI & TECH
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