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Primer and method for molecular identification of new whitebait species in Taihu Lake

A technology of molecular identification and whitebait, which is applied in biochemical equipment and methods, measurement/testing of microorganisms, DNA/RNA fragments, etc., can solve the problems of inaccurate PCR product results and unstable primer amplification, etc., and achieve The PCR product is accurate and reliable, the result is stable, and the effect of stable amplification

Inactive Publication Date: 2014-11-19
HUAINAN NORMAL UNIV
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Problems solved by technology

[0010] However, in the later research of the inventor, it was found that in the process of using the above primers to detect the mitochondrial COⅡ gene of whitebait to construct a biological tree, the amplification of the primers was not very stable, and the results of PCR products were not accurate enough. To accurately identify the species of whitebait, Need to redesign a primer that can stably amplify

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  • Primer and method for molecular identification of new whitebait species in Taihu Lake
  • Primer and method for molecular identification of new whitebait species in Taihu Lake
  • Primer and method for molecular identification of new whitebait species in Taihu Lake

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Embodiment 1

[0042] Such as figure 1 Shown, the method step of a kind of Taihu Lake new whitebait species molecular identification of the present invention is:

[0043] Step 1. Construct a standard stream biological tree

[0044] The complete and fresh individual samples of Taihu Newbait were obtained, compared with the indoor samples of animal specimens of Huainan Normal University, and it was initially confirmed as a sample of Taihu Newbait, and then based on the morphological characteristics of Taihu Newbaba (number and position of tongue and teeth, fins) position and number of fin rays), use dissecting mirror and microscope to carry out morphological identification, confirm the species, and use it as the standard specimen of new whitebait in Taihu Lake. The laboratory takes the fresh muscle tissue of the specimen, uses SDS / protease to lyse, and extracts with phenol chloroform Total DNA was detected by agarose gel electrophoresis, and the total DNA content was determined by spectrophot...

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Abstract

The invention discloses a primer and a method for molecular identification of new whitebait species in Taihu Lake, which belong to the molecular biology DNA mark field. The primer sequence is ND5L (5'-CTTGGTGCAAATCCAAGCAGGAAC-3), ND5R(5'-GCTTACTCGTGGCCTGAGCCGA-3'); The identification method comprises the following steps: 1)using the primer pair for PCR amplification on total DNA of a new whitebait experiment specimen in Taihu Lake, 2)purifying the PCR products, copying and cloning, and sequencing to obtain a ND5 gene complete sequence; 3)comparing the experiment specimen with the ND5 gene complete sequence of related species to construct a standard flow biological genealogical tree; 4)acquiring the complete sequence of gene of a fish sample to be detected by the above method, adding the sequence in the step 3) to reconstruct a detection flow biological genealogical tree; and 5)analyzing and comparing. The primer and method are used for accurately identifying and classifying species with non morphology, and provide scientific identification basis for artificial culture and hybridization of species as well as genetic research.

Description

technical field [0001] The invention belongs to the technical field of DNA markers of molecular biology, and more specifically relates to a primer and a method for molecular identification of new whitebait species in Taihu Lake. Background technique [0002] Whitebait (icefishes) is an important economic fish in my country. China has 15 species of whitebait among the 17 species in the world, of which 6 species are endemic to China. Taihu new whitebait (Neosalanx taihuensi) is a relatively common whitebait, endemic to China, belonging to the genus Neosalanx (Salangidae) of the family Salangidae. [0003] Whitebait has high nutritional value and economic value: every 100 grams of fresh whitebait edible part contains 8.2 grams of protein, 0.3 grams of fat, 1.4 grams of carbohydrates, and 258 mg of calcium. Whole fish can be used as medicine. Its meat is sweet and flat. It has the effects of nourishing kidney, invigorating stomach and tonifying deficiency, benefiting lung and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 张际峰周杰王建超
Owner HUAINAN NORMAL UNIV
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