Constructing method of standard sample in Alexandrium catenella (A.catenella) fluorescence quantitative polymerase chain reaction

Abstract

The invention relates to a constructing method of a standard sample in Alexandrium catenella (A.catenella) fluorescence quantitative polymerase chain reaction. A recombinant plasmid is constructed by extracting DNA of A.catenella ACDH01, amplifying 18S rDNA of A.catenella ACDH01 to a 28S rDNA gene segment through polymerase chain reaction, and connecting an amplified product with a T carrier; the confirmed recombinant plasmid is diluted into a polymerase chain reaction standard sample in a gradient way; real-time polymerase chain reaction is carried out by using 5.8S-b5' and 5.8S-b3' as primers and a recombinant plasmid standard sample as a template; and a standard curve is prepared according to a clinical circulating number in the reaction and the log value of the gradient concentration of the recombinant plasmid standard sample. The recombinant plasmid real-time polymerase chain reaction standard sample with 18S rDNA-28S rDNA gene segments of the A.catenella ACDH01 is established; and the real-time polymerase chain reaction detection is accurately, efficiently and rapidly carried out on A.catenella ACDH01 to be tested by adopting a pair of universal primers.

Description

technical field [0001] The invention relates to a construction method of axandrium fluorescent quantitative polymerase chain reaction standard product, which is used for molecular biology detection of marine red tide algae and belongs to the technical field of microbial molecules. Background technique [0002] Since 1972 in my country, the economic loss caused by red tides has reached more than 1 billion yuan per year, and some extremely large red tides can affect thousands of square kilometers of sea area at a time, causing economic losses of several hundred million yuan. Especially in recent years, the frequent occurrence of red tides has caused huge losses to the country. Therefore, the detection of red tide algae is particularly important. The rapid, sensitive and accurate detection of red tide algae can play a role in early warning of red tide outbreaks. [0003] At present, toxic red tide algae are mainly detected by morphological classification methods, immunology, a...

AI Technical Summary

Problems solved by technology

In the past, using DNA or PCR products as standard products had the disadvantage of instability, which could easily lead to inaccurate results.

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