Spider mitochondria genome complete sequence amplification primer and amplification method

A mitochondrial genome and amplification primer technology, applied in recombinant DNA technology, DNA preparation, DNA/RNA fragments, etc., can solve problems such as unfinished sequencing work, and achieve the effect of saving the cost of reagents and reducing the number of reagents

Active Publication Date: 2015-04-22
CHINA JILIANG UNIV
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Problems solved by technology

At present, only 13 species of spiders have been sequenced for the complete mitochondrial genome, but there are many kinds of s

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  • Spider mitochondria genome complete sequence amplification primer and amplification method

Examples

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Embodiment 1

[0035] (1) Collection, identification and preservation of spider samples

[0036] The species of spiders used in the examples were all collected from the farmlands of Yuyao City or Tongxiang County, Zhejiang Province by trap method. The collected specimens were brought back to the laboratory for information registration, and all were identified with a stereo microscope to determine the species. The identified spider samples were soaked in 100% alcohol and stored in a 4°C refrigerator for later use. The species of spiders collected include: Pseudomonas tarantula, Triflower spider, Golden spider, Cone-bellied octopus, Ornate octopus, Brown new orb spider, Maiwa spider, black fly tiger, big-bellied garden spider, cat jumping spider.

[0037] (2) Extraction of Total DNA from Spider Genome

[0038] DNA extraction kits were used to extract the total genomic DNA of different species of spiders. To avoid contamination, the head and abdomen were discarded, and DNA was extra...

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Abstract

The invention discloses a spider mitochondria genome complete sequence amplification primer and an amplification method. The method comprises the steps of extracting a general DNA of the spider genome, designing a mitochondria genome complete sequence universal amplification primer, and amplifying and sequencing the long PCR (L-PCR) of the DNA of the mitochondria genome. The spider mitochondria genome complete sequence amplification primer consists of 6 pairs of single strain oligonucleotides. Multiple spider mitochondria genome complete sequences can be quickly and efficiently amplified by adopting the universal amplification primer, and a material basis is established to spider species identification, population diversity analysis, genetic resource protection and utilization, systematic inheritance evolution and other researches.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to a primer for amplifying the complete sequence of a spider mitochondrial genome and an amplification method. Background technique [0002] Mitochondria is an important organelle in cells, and its genomic DNA is in a covalently closed circular shape with autonomous replication, transcription and translation capabilities. The mitochondrial genome generally consists of 13 protein genes, 22 tRNA genes, 2 rRNA genes (12S rRNA and 16S rRNA), a total of 37 coding genes and a non-coding region (control region). Compared with nuclear DNA, mitochondrial genomic DNA has the advantages of maternal inheritance, large copy number, low recombination rate, and fast evolution. tool. For example, some genes on mitochondria (such as COI, CytB, 16S rRNA, etc.) can be used as important genetic molecular markers in the study of species classification and identification. The co...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12N15/10
Inventor 王正亮俞晓平李超
Owner CHINA JILIANG UNIV
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