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Primer for enlarging species specificity of S. calospora

A kind of specific and giant spore technology, applied in the direction of biochemical equipment and methods, measurement/testing of microorganisms, DNA/RNA fragments, etc., can solve the problems that cannot be directly amplified

Inactive Publication Date: 2010-10-20
南通国邦科技发展有限公司
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Problems solved by technology

[0003] The present invention aims to solve the problem that existing primers cannot directly amplify the partial sequence of the 25S rDNA D1 / D2 variable region with the genetic characteristics of Scutellospora calospora from mixed DNA samples, and provides a Species-specific primers used to amplify Megaspora glabrata

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  • Primer for enlarging species specificity of S. calospora

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specific Embodiment approach 1

[0009] Specific embodiment 1: In this embodiment, the species-specific primer used to amplify Scutellaria sp. is 5'-TTAAAGCCATTACGTCAGCG-3'.

specific Embodiment approach 2

[0010] Specific embodiment two: This embodiment is used to amplify the species-specific primer 5'-TTAAAGCCATTACGTCAGCG-3' used to amplify Scutellaria sp. as a reverse primer.

[0011] The forward primer LR1 (5'-GCATATCAATAAGCGGAGGA-3') was paired with the reverse primer of this embodiment. Scutellospora calospora DNA-containing mixed samples, Scutellospora calospora DNA samples and mixed samples without Scutellospora calospora DNA were used as templates to perform primer S.cal specificity verification tests. The PCR amplification system and reaction conditions are as follows:

[0012] The amplification reaction system consists of 20μL of 2.0μL DNA template, 1.6μL of dNTP with a concentration of 2.5mmol / L, 2.0μL of 10× buffer (without Mg 2+ ), 2.0μL of MgCl at a concentration of 25mmol / L 2 , 2.0μL of primers with a concentration of 10pmol / L, 0.2μL of Taq enzyme (DNA polymerase) with a concentration of 5U / μL and the remainder of redistilled water; amplification reaction conditions: p...

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Abstract

Disclosed is a species specificity primer used for amplifying Scutellospora calospora, which relates to a specific primer used for amplifying the Scutellospora calospora. The primer solves the problem that the existing primer can not directly amplify the sequence at the 25S rDNA D1 / D2 variable area part having hereditary properties of Scutellospora calospora from a mixed DNA sample. The species specificity primer for amplifying the Scutellospora calospora is 5`-TTAAAGCCATTACGTCAGCG-3`. The matching of the primer of the invention and an LR1 primer in a general matched primer can accurately amplify target fragments of Scutellospora calospora.

Description

Technical field [0001] The present invention relates to a specific primer for amplifying Megacystis scutellariae. Background technique [0002] Scutellospora calospora (Scutellospora calospora) is a kind of arbuscular mycohizal (AM) fungus, because mycorrhizal includes endophytic, exophytic, endophytic mycorrhiza, etc., exophytic fungi can be cultivated, while AM Fungus is a kind of endophytic mycorrhizal fungus, it is not cultivable. Without the necessary plant root symbiosis, the life history cannot be completed, and the reproduction and survival cannot be continued, that is, it cannot be purely cultivated. AM fungi cannot be cultured purely. Although a wet sieve decantation method can be used to obtain a single AM ​​fungus spore from plant rhizosphere soil, and then DNA extraction of a single AM ​​fungus spore can be amplified by using universal eukaryotic primers. The sequence can be known by techniques such as sequencing, but if it is in a mixed DNA sample where multiple b...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12Q1/04C12N15/11
Inventor 蔡柏岩接伟光葛菁萍王雪阎秀峰
Owner 南通国邦科技发展有限公司
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