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Oil-tea SSR molecular marker

A molecular marker, Camellia oleifera technology, which is applied in the development of Camellia oleifera microsatellite genetic markers, can solve the problems of little research on the development and utilization of Camellia oleifera microsatellite molecular markers, and achieves high reliability and good repeatability.

Active Publication Date: 2014-03-26
JIANGXI ACAD OF FORESTRY
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Problems solved by technology

However, so far, there has been little research on the development and utilization of camellia oleifera microsatellite molecular markers, and there are only a few domestic reports on the use of tea tree microsatellite marker primers to randomly amplify the camellia oleifera genome (Liu Bing et al., Establishment and optimization of Camellia oleifera SSR-PCR reaction system. Journal of Anhui Agricultural University ,2011,38(6):858-862), in addition to Wen Q et al.Development of polymorphic microsatellite markers in Camellia chekiangoleosa(Theaceae),American Journal of Botany,2012,5(1):e203- e205.) Except for the 18 pairs of SSR markers developed by using the EST sequence of Camellia oleifera species, Zhejiang safflower camellia, there is no report on the development of SSR markers of Camellia oleifera by using the DNA sequence of camellia oleifera

Method used

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  • Oil-tea SSR molecular marker
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  • Oil-tea SSR molecular marker

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Embodiment 1

[0061] 1. Preparation method of camellia oleifera SSR molecular marker

[0062] 1.1 Acquisition and pretreatment of Camellia oleifera DNA sequence

[0063] Using the method of our research group (refer to Jiang C et al. Efficient extraction of RNA from various Camellia species rich in secondary metabolites for deep transcriptome sequencing and gene expression analysis. African Journal of Biotechnology, 2011, 10(74): 16769-16773. ) to extract Camellia oleifera DNA and RNA, construct genome and transcriptome libraries respectively, and obtain genome and EST sequences through each 1 / 4 454GS FLX high-throughput sequencing reaction. The EST sequence was spliced ​​by CAP3 after removing low-quality sequences by SeqClean and Lucy software, and the genome sequence obtained by sequencing was processed and spliced ​​by the Newbler software that comes with the 454 sequencer.

[0064] 1.2 Screening of SSR loci in Camellia oleifera DNA sequence

[0065] The SSR sites in each DNA sequence...

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Abstract

The invention discloses an oil-tea SSR molecular marker which comprises acquisition and pre-treatment of an oil-tea DNA sequence, retrieval of SSR loci in the DNA sequence, design and synthesis of an SSR primer, establishment of an SSR marker system, and screening of a polymorphic SSR marker. Oil-tea genomic sequences obtained by 454 sequencing and eligible sequences containing simple sequence repeat units in an EST sequence are retrieved and screened to obtain twelve polymorphic SSR markers: CO_gSSR11, CO_gSSR12, CO_gSSR14, CO_gSSR16, CO_gSSR19, CO_gSSR20, CO_eSSR01, CO_eSSR04, CO_eSSR16, CO_eSSR40, CO_eSSR44 and CO_eSSR48. The oil-tea SSR molecular marker disclosed by the invention is suitable for genetic evaluation, germplasm identification and molecular marker assisted breeding research of oil-tea germplasm resources, has the advantages of good repeatability and high reliability, and is an effective molecular marker.

Description

technical field [0001] The invention relates to a DNA molecular marker technology, in particular to the development technology of microsatellite genetic markers for camellia oleifera, which is suitable for genetic diversity research, germplasm identification and kinship analysis of camellia oleifera. Background technique [0002] Microsatellite (Microsatellite) refers to a DNA sequence that is repeated in series with a few nucleotides (mostly 2-4) widely distributed in the genome, which is called simple sequence repeats (Simple sequence repeats) , SSR), is a type of DNA sequence with a relatively high mutation rate in the genome. In recent years, SSR has become the most popular genetic marker. As a co-dominant marker with high detection efficiency, SSR markers combine the advantages of other molecular markers, such as typical co-dominant and multi-allelic genes, with high polymorphism, and can accurately distinguish individuals with very similar kinship; The amount of DNA ...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68
Inventor 温强刘丽婷徐立安徐林初周文才叶金山朱恒黄敏仁
Owner JIANGXI ACAD OF FORESTRY
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