Universal primers for amplifying long fragment of Plecoptera insect mitochondrial genome

A universal primer, mitochondrial technology, applied in the direction of DNA/RNA fragment, recombinant DNA technology, microorganism determination/inspection, etc., can solve the problems of long time, high template purity requirements, lack of specificity, etc., to shorten the experimental period, shorten the The effect of obtaining cycle and reducing experiment cost

Inactive Publication Date: 2016-11-09
YANGZHOU UNIV
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  • Abstract
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Problems solved by technology

This method uses a large number of primers, low amplification efficiency, high requirements for template purity, long time-consuming, and lack of sufficient specificity, which will hinder the rapid accumulation of molecular data of insect m

Method used

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  • Universal primers for amplifying long fragment of Plecoptera insect mitochondrial genome
  • Universal primers for amplifying long fragment of Plecoptera insect mitochondrial genome
  • Universal primers for amplifying long fragment of Plecoptera insect mitochondrial genome

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Embodiment Construction

[0013] The amplification primer screening method for long fragments of the Anoptera mitochondrial genome of the present invention is divided into two steps: 1. Based on the mitochondrial genome sequences of insects of different orders measured in Genbank, relatively conservative sequence fragments are found by comparison Design primers and introduce degenerate sites to increase the generality of the primers; 2. Amplify the synthetic primers to the mitochondrial genome of the order Anoptera, and test their generality and amplification efficiency in the order; 3. Many pairs of primers have been designed in the process, and the verification of the experimental results shows that the primers of the present invention are the best.

[0014] Below in conjunction with example the invention is further described:

[0015] 1. Sample preparation: The samples are all insects of different families of the order Anoptera stored in the specimen room of the Institute of Applied Entomology, Yang...

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Abstract

The invention discloses universal primers for amplifying a long fragment of a whole Plecoptera insect mitochondrial genome. The universal primers comprise two pairs of primers with the sequence shown as SEQ ID NO.1-SEQ ID NO.4. With the adoption of the universal primers, long fragments of mitochondrial genomes of various Plecoptera species can be effectively amplified, the acquisition cycle of molecular data can be shortened, and an important tool is provided for phylogenic research of Plecoptera insects in China.

Description

technical field [0001] The invention relates to two pairs of universal primers for amplifying the long fragments of mitochondrial genomes of insects of the order Anoptera. Background technique [0002] Plecoptera, also known as stoneflies, are widely distributed in various types of waters and are important water quality monitoring indicator organisms. Strengthening the taxonomic research of this group can better provide water quality environment monitoring. Serve. [0003] From the point of view of systematics and evolution, Chiroptera is a key group. Although the Cereoptera phylogeny has been extensively studied, it still needs to be proved by other more characters. The data obtained by molecular means will be more helpful in inferring the genetic relationship among the families of Chiroptera and the genetic relationship with other orders. An in-depth understanding of the origin, evolution, and phylogenetic relationship of Euoptera insects is of great significance for re...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12N15/10
CPCC12N15/11C12Q1/6806
Inventor 杜予州陈志腾
Owner YANGZHOU UNIV
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