Method for sequencing partial DNA of mitochondria ND5 gene of brachionus rotifera and authenticating brachionus rotifera

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

Inactive Publication Date: 2013-05-08
ANHUI NORMAL UNIV
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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...

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[0014] The biological materials used in this method for identifying rotifers of the genus Brachionus are Brachionus rubens, Brachionus quadridentatus, Brachionus falcatus, Brachionus diversicornis, tail process Brachionus caudatus, Brachionus caudatus, Brachionus caudatus, single individuals of each type of rotifer were picked and cultured in monoclonal manner, among them, Brachionus caudatus and Brachionus square used the formula of Gilbert (1963), and the others used the formula 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 cultured with HB-4 culture medium and was in the exponential growth phase of Scenedesmus oblique. When the number of single rotifers reached When there is a certain amount, the rotifers are filtered out with a 25 μm pore size filter, rinsed with sterile distilled water, collected in Eppendof tubes, and stored in a -20°...

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Abstract

The invention relates to methods for sequencing partial DNA of a mitochondria ND5 gene of brachionus rotifera and authenticating the brachionus rotifera. The sequencing method comprises the following steps of: extracting the total DNA of the brachionus rotifera, amplifying and sequencing polymerase chain reaction (PCR) primer and analyzing system generation. The authenticating method comprises the following step of: authenticating the DNA of the mitochondria ND5 of the brachionus rotifera. The PCR primer designed in the method can amplify the DNA of the mitochondria ND5 of the brachionus rotifera; a sequencing result can be used for precisely authenticating the brachionus rubens and the like; the difficulty in authenticating caused by morphology and an ultra microstructure is solved; and in a process of cultivating the rotarian in large scale, the probability that the number of rotaria is reduced or massive rotaria are dead caused by population competition because of mixed populations is reduced.

Description

technical field [0001] The invention belongs to the field of biological technology, and in particular relates to a partial DNA sequencing of the mitochondrial ND5 gene of the rotifer Brachionus and a method for identifying the rotifer Brachionus. 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 ...

Claims

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Application Information

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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
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