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PCR (Polymerase Chain Reaction) primer group for distinguishing five types of shellfish bait microalgae

A technology of bait microalgae and primer set, applied in the direction of microbe-based methods, microbes, microbe measurement/inspection, etc., can solve problems such as easy cross-amplification of primers, deviation of detection results, and influence on production and application

Inactive Publication Date: 2012-09-19
NINGBO UNIV
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Problems solved by technology

However, due to the high genetic similarity of the five bait microalgae, the designed primers are prone to cross-amplification, which will cause deviations in the detection results and affect production applications.

Method used

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  • PCR (Polymerase Chain Reaction) primer group for distinguishing five types of shellfish bait microalgae
  • PCR (Polymerase Chain Reaction) primer group for distinguishing five types of shellfish bait microalgae
  • PCR (Polymerase Chain Reaction) primer group for distinguishing five types of shellfish bait microalgae

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Embodiment Construction

[0025] The present invention will be further described in detail below in conjunction with relevant experimental data.

[0026] 1. Primer design:

[0027] The present invention clones and sequences the full-length (about 1700bp) 18S rDNA genes of five common bait microalgae between different genera, and uses the DNA sequence alignment and analysis software MEGA 5.0 and DNAMAN 6.0 to analyze and compare. The results show that: five species The difference percentage of 18S rDNA sequence in microalgae was 8.32%~20.42%, and the average difference was 11.14%.

[0028] Through the analysis, it was found that the pairwise difference regions of the 18S rDNA of the five microalgae are not pure and absolute differences, and there are often multiple homologous sites in the difference regions, and these homologous sites have a length of about 20 bp The design of the primers caused great interference, which is mainly reflected in the fact that the primers have a certain annexation effect ...

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Abstract

The invention relates to a PCR (Polymerase Chain Reaction) primer group for distinguishing five types of shellfish bait microalgae, and the PCR primer group is used for identifying Chaetoceros of Bacillariophyta, Isochrysis galbana and Pavlova viridis of Chrysophyta and Platymonas helgolandica and Nannochloris oculata of Chlorophyta, and nucleotide sequences of primers are respectively SEQ ID NO: 1-10. According to the invention, specificity primers of five types of bait microalgae can be provided, a PCR technology is combined to carry out in vitro amplification for 18S r DNA (Deoxyribonucleic Acid) of microalgae, and an amplification result can be analyzed through an AGE (Agarose Gel Electrophoresis) technology. Through the amplification result, the primers and templates can be better combined, and an amplification reaction can be triggered, so that whether the five types of shellfish bait microalgae exist in a sample or not can be quickly sensitively distinguished, detected and identified.

Description

technical field [0001] The invention belongs to the technical field of microalgae identification, in particular to a PCR primer set for distinguishing five kinds of shellfish bait microalgae, that is, for quickly and sensitively distinguishing and detecting Chaetoceros calcitrans and Chrysophyta Specific PCR primer sets for Isochrysis galbana, Pavlova viridis, Platymonas helgolandica and Nannochloropsis oculata of the Chlorophyta. Background technique [0002] Marine microalgae are the earliest biological groups that appeared on the earth. They are small in size, large in number, and fast in reproduction. As the main primary producers in the marine ecosystem, they play an extremely important role in the material cycle and energy flow of the marine ecosystem. important role. Many marine microalgae are rich in a variety of polyunsaturated fatty acids (PUFA), which have extremely high nutritional value for coastal shellfish. In addition, they have the advantages of rapid growt...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11C12R1/89
Inventor 朱鹏周海波徐继林严小军
Owner NINGBO UNIV
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