Primer pair, castanopsis hystrix SSR3 (Simple Sequence Repeat 3) marker and preparation method and application thereof

A primer pair and red cone technology, which is applied in biochemical equipment and methods, DNA preparation, microbial measurement/inspection, etc., can solve the problems of poor stability, inability to directly apply molecular marker-assisted breeding and QTL mapping, etc.

Inactive Publication Date: 2014-04-23
GUANGXI FORESTRY RES INST
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  • Abstract
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Problems solved by technology

[0004] Although scholars at home and abroad have used multiple markers such as ISSR and RAPD to study the genetic diversity of red cone populations, the genetic structure of red cone populations, and the genetic relationship and distance between red cones of different provenances, the two markers of ISSR and RAPD All are dominant markers with poor stability and cannot be directly applied to molecular marker-assisted breeding and QTL mapping

Method used

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  • Primer pair, castanopsis hystrix SSR3 (Simple Sequence Repeat 3) marker and preparation method and application thereof
  • Primer pair, castanopsis hystrix SSR3 (Simple Sequence Repeat 3) marker and preparation method and application thereof
  • Primer pair, castanopsis hystrix SSR3 (Simple Sequence Repeat 3) marker and preparation method and application thereof

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Embodiment

[0028] Extraction and enzyme digestion of total DNA of red cone

[0029] Select the young leaves of Red Cone, and use CTAB method to extract the nuclear genome; digest with EcoRV at 37°C for 3 hours, and inactivate the endonuclease at 65°C for 20 minutes; the reaction system is 20 μL, including 2.0 μL of 10×Buffer, 1.0 μL EcoRV endonuclease, 2 μL of DNA, 15.0 μL of sterilized double distilled water;

[0030] Connection of connector

[0031] Connect the red cone genomic DNA digested by EcoRV in step to the upper and lower linkers (upper linker sequence GTAATACGACTCACTATAGGGCACGCGTGGTCGACGGCCCGGGCTGGT, sequence listing SEQ.ID.No.4; lower linker sequence ACCAGCCC-NH2, sequence listing SEQ.ID.No.5); The 60 μL ligation system contains 9 μL of enzyme-cut red cone genomic DNA, and 100 pmol of the upper and lower adapters; the ligation reaction is carried out under the condition of 1 times T4 ligase buffer, and ligated overnight at 16°C; the ligated mixture is reacted at 65°C for 2...

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Abstract

The invention discloses a primer pair, a castanopsis hystrix SSR3 (Simple Sequence Repeat 3) marker and a preparation method and application thereof. According to the preparation method, the total DNA (Deoxyribonucleic Acid) of castanopsis hystrix is subjected to PCR (Polymerase Chain Reaction) amplification by using the specific primer pair HZ3, thereby obtaining the SSR3 marker. The method is simple and feasible and is simple and convenient in operation; shown by tests which are carried out in members of two natural castanopsis hystrix populations and among the members, the obtained SSR has relatively high polymorphism. The castanopsis hystrix SSR3 marker is a co-dominant marker; compared with other molecular markers, the SSR marker is good in repeatability and high in reliability, can be applied to the genetic diversity evaluation of castanopsis hystrix, the genetic map construction of castanopsis hystrix, the research on the spatial distribution pattern of the genetic variation of castanopsis hystrix population and the origination and evolution of castanopsis hystrix and the standardization of target genes, and can also be applied to the molecular marker assisted breeding and QTL (Quantitative Trait Locus) research of castanopsis hystrix.

Description

technical field [0001] The invention belongs to the technical field of molecular markers in biological sciences, and in particular relates to a red cone SSR3 marker, a pair of primers and a preparation method and application thereof. Background technique [0002] Castanopsis hystrix is ​​an evergreen tree belonging to the genus Castanopsis of the Fagaceae family. It is an important native broad-leaved, high-quality and precious timber tree species in South China. It grows fast, adapts widely, and has high benefits. It can be used for construction, shipbuilding, and high-end furniture. It has strong germination power, dense branches and leaves, is more resistant to shade, and has good mixed growth performance. It can be planted in pure forests or mixed forests. It is a mixed forest of coniferous species. One of the most ideal companion tree species; the red cone has strong adaptability to the habitat, and is located in the arbor layer in the community. Because red cone wood ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12N15/10C12Q1/68
Inventor 蒋燚刘海龙朱积余黄荣林姜英何应会
Owner GUANGXI FORESTRY RES INST
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