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Sequencing method for part of DNA (deoxyribonucleic acid) of mitochondrial ND4 genes of Brachionus rotifer and identification method of Brachionus rotifer

A technology of DNA sequencing, DNA sequence, applied in the field of biology

Inactive Publication Date: 2015-03-11
ANHUI NORMAL UNIV
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  • Description
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  • Application Information

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Problems solved by technology

The identification of rotifers by molecular biology solves the identification difficulties of using morphology and ultrastructure, so that in the process of large-scale cultivation of rotifers, the population competition caused by population mixing can be reduced, resulting in reduced production of rotifers or even possibility of mass death

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  • Sequencing method for part of DNA (deoxyribonucleic acid) of mitochondrial ND4 genes of Brachionus rotifer and identification method of Brachionus rotifer
  • Sequencing method for part of DNA (deoxyribonucleic acid) of mitochondrial ND4 genes of Brachionus rotifer and identification method of Brachionus rotifer
  • Sequencing method for part of DNA (deoxyribonucleic acid) of mitochondrial ND4 genes of Brachionus rotifer and identification method of Brachionus rotifer

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Embodiment Construction

[0014] The biological materials used in this method for identifying the genus Brachionus rotifers are Brachionus redus, Brachionus scissors, Brachiona square, Brachiona schizopoda, Brachiona falciparum, Brachionus tendigitata For rotifers, Brachionus occipitalis, and Brachionus rectangularis, single individuals of each species were selected for monoclonal culture, among which Brachionus redus and Rotifer squares used the formula of Gilbert (1963), and the rest The original local water samples were filtered and used as culture medium. The temperature was basically the same as the ambient temperature at the time of collection. The food used was Scenedesmus obliquus in the exponential growth phase cultured with HB-4 culture medium. When a single clone rotifer individual When the number reached a certain amount, the rotifers were filtered out with a 25 μm pore size filter, washed with sterile distilled water, collected in Eppendof tubes, and stored in a -20°C refrigerator.

[0015...

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Abstract

The invention relates to a sequencing method for a part of DNA (deoxyribonucleic acid) of mitochondrial ND4 genes of Brachionus rotifer and an identification method of the Brachionus rotifer. The sequencing method comprises the following steps of: extracting total DNA (deoxyribonucleic acid) of the Brachionus rotifer, performing PCR (polymerase chain reaction) primer amplification, determining a sequence and performing phylogenetic analysis; and according to the identification method, mitochondrial ND4 DNA of the Brachionus rotifer is used for identification. A designed PCR primer in the sequencing method can be used for amplifying the mitochondrial ND4 DNA of Brachionus rubens and the like, a sequencing result can be used for precisely identifying the Brachionus rubens and the like, the difficulty in identification by utilizing morphology and an ultra-micro structure can be solved, and then the possibility of drop in production and even mass deaths of the rotifers caused by population competition in mixed populations during the large-scale culture of the rotifers can be further reduced.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for partial DNA sequencing of the mitochondrial ND4 gene of Brachionus rotifers and identification thereof. Background technique [0002] Early taxonomic and systematic studies of rotifers were mainly based on morphological data. For example, Wallace (1989) conducted cluster analysis using eight morphological features and found that Rotifera, Eutatoria, and Monogonota were monophyletic, and clustered using nine morphological features. Class analysis found that the genus Notholca was divided into six clades. Melone (1998) obtained a parsimony clustering tree through matrix analysis of 60 morphological features of 1 acanthocephala, 6 rotifers and 2 outgroups (planarians and gnathogastrics), and found that the Dinestidate ( Digononta) is a paraphyletic group, but the monophyletic of rotifers cannot be determined. The phylogenetic tree is different from the phylog...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 程双怀张逸夏梦宁刘应龙卢燕梅闫锦锦张宁
Owner ANHUI NORMAL UNIV