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SNP locus multicolor fluorescence detection primer and kit used for identifying eucalyptus clones, detection method and applications

A technology of fluorescence detection and identification method, which is applied in the field of multicolor fluorescence detection primers for identification of SNP sites of eucalyptus clones, can solve the problems of no SNP detection system, etc., and achieve protection of rights and interests, high site polymorphism, and convenient operation Effect

Active Publication Date: 2019-08-06
RES INST OF TROPICAL FORESTRY CHINESE ACAD OF FORESTRY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, although some DNA fingerprint analysis work has been carried out on Eucalyptus clones, there is still no simple and easy SNP detection system

Method used

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  • SNP locus multicolor fluorescence detection primer and kit used for identifying eucalyptus clones, detection method and applications
  • SNP locus multicolor fluorescence detection primer and kit used for identifying eucalyptus clones, detection method and applications
  • SNP locus multicolor fluorescence detection primer and kit used for identifying eucalyptus clones, detection method and applications

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Experimental program
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Embodiment 1

[0033] (1) Eucalyptus clone DNA extraction

[0034] The leaves of eucalyptus clones (Table 2) were collected, and genomic DNA was extracted by CTAB method, and frozen after detection for future use.

[0035] Table 2 58 Eucalyptus clones

[0036]

[0037] Weigh 0.3g of 58 Eucalyptus clones (Table 2) leaf samples, grind them in liquid nitrogen, transfer to a 2mL centrifuge tube, add 1mL CTAB extract, incubate at 60-65°C for 45-60min, and shake once every 10min. Take out the sample tube, centrifuge at 12000rpm at 4°C for 10min, and take the supernatant. Add an equal volume of chloroform and isoamyl alcohol mixture (chloroform:isoamyl alcohol=24:1, volume ratio), seal, shake well; centrifuge at 4°C, 12000rpm for 10min, and take the supernatant. Add an equal volume of chloroform and isoamyl alcohol mixture (chloroform:isoamyl alcohol=24:1, volume ratio), centrifuge at 12000rpm for 10min, and take the supernatant. Add refrigerated, 2 / 3 volume isopropanol to the supernatant, sh...

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Abstract

The invention discloses a SNP locus multicolor fluorescence detection primer and kit used for identifying eucalyptus clones, a detection method and applications. SNP markers with high polymorphism arescreened in a eucalyptus genome; multicolor fluorescence multiple detection systems of 21 loci are optimized and established; and typing results have the characteristics of being easy in interpretation of banding patterns, high in locus polymorphism and easy in fluorescence distinguishing. The method has the advantages of being high in experimental efficiency, accurate in detection and convenientin operation; important technical support can be provided for rapidly performing germplasm resource genetic evaluation, molecular marker-assisted breeding and construction and identification of clonal molecular fingerprints on eucalyptus in the future; the molecular fingerprints of 58 eucalyptus clones commonly used in China are constructed, identification can be successfully performed on the above clones, so that the workload of eucalyptus clone identification can be greatly reduced; and the method can effectively discriminate counterfeit and disordered clones, so that the rights and interests of breeding cultivation people and forest culture and management planters can be practically guaranteed.

Description

technical field [0001] The invention belongs to the technical field of molecular markers, and in particular relates to SNP site multicolor fluorescence detection primers, kits, detection methods and applications for identifying eucalyptus clones. Background technique [0002] Eucalyptus is the general term for species of Eucalyptus, Angophora and Corymbia in the family Myrtaceae. It has the characteristics of rich genetic diversity, rapid growth and strong adaptability. It is widely planted in subtropical and subtropical regions, and plays an important role in alleviating the tense situation of timber supply and demand in my country. [0003] Eucalyptus in production is mainly afforested in the form of clone tissue culture. However, with the increase in the number of tissue culture propagation, the expansion of the planting area and the expansion of the distribution market, the problem of eucalyptus clones being wrongly recorded and shoddy is becoming more and more serious. ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11C12Q1/6858
CPCC12Q1/6895C12Q1/6858C12Q2600/13C12Q2600/156C12Q2521/319C12Q2521/525C12Q2531/113Y02A50/30
Inventor 周长品李发根翁启杰李昌荣甘四明
Owner RES INST OF TROPICAL FORESTRY CHINESE ACAD OF FORESTRY
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