Primers for detecting SSR (simple sequence repeat) marks of prunus pseudocerasus

A technology for detecting primers and cherries, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., and can solve the problem of insufficient number of molecular markers in cherries

Active Publication Date: 2016-02-17
ZHEJIANG NORMAL UNIVERSITY
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Problems solved by technology

[0004] The technical problem to be solved in the present invention is to overcome the shortcomings of insufficient number of cherry molecular markers in the prior art, provide a set of primers for detecting SSR markers of plants of the genus Cherry, and increase the number of molecular markers of the genus Cherry to be applied to future breeding work middle

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  • Primers for detecting SSR (simple sequence repeat) marks of prunus pseudocerasus
  • Primers for detecting SSR (simple sequence repeat) marks of prunus pseudocerasus
  • Primers for detecting SSR (simple sequence repeat) marks of prunus pseudocerasus

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Embodiment 1

[0056] In the present invention, the specific method of using the SSR marker developed by the transcriptome of Chinese cherry "Short handle cherry" is:

[0057] 1. DNA extraction

[0058] (1) Preparation of DNA extraction solution: 2% CTAB, 0.1 MTris, 20 mM EDTA, 1.4 M NaCl, pH 8.0; DNA dissolution buffer (ie TE buffer): 10 mM Tris; 1 mM EDTA; pH=8.0.

[0059] (2) Chinese cherry material is processed as follows:

[0060] ① Add 4 mL of DNA extraction solution and 80 μL of β-mercaptoethanol to a 10 mL centrifuge tube, and place it in a 65°C water bath to preheat for 10 minutes;

[0061] ②Take 1g of fresh leaves, put a little PVP in the mortar, add liquid nitrogen to grind thoroughly; transfer the ground leaves to a centrifuge tube containing the extract, mix them well and put them in a water bath at 65°C for 30 minutes. Shake once every 10 minutes to fully lyse the leaf cells;

[0062] ③After the water bath, take out the centrifuge tube and cool it to room temperature; add an...

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Abstract

The invention provides primers for detecting SSR (simple sequence repeat) marks of prunus pseudocerasus. The group of primers comprises nine primer pairs which can be used simultaneously in the detection process, wherein the sequences of the nine primer pairs are shown in SEQ ID NO:1 to SEQ ID NO:18. The nine SSR marks screened out have polymorphism in species or varieties of prunus pseudocerasus, genetic diversity analysis performed in 22 species or varieties of prunus pseudocerasus basically coincides with classification common sense of cerasus plants, the SSR marks are new marks existing stably, and the nine marks provided in the invention can be directly applied to more prunus pseudocerasus or even transferred to prunus avium for genetic diversity analysis of germplasm resources, construction of genetic maps and molecular mark assisted breeding.

Description

technical field [0001] The invention relates to the field of molecular biology DNA marker technology and application, in particular to primers for detecting cherry SSR markers. Background technique [0002] Simple Sequence Repeat (SSR), also known as tandem repeat sequence or microsatellite marker, is a co-dominant marker with the advantages of high polymorphism, good repeatability, and easy detection, and is widely used in germplasm resources Genetic relationship identification, genetic map construction, functional gene screening and molecular marker-assisted breeding. There are usually three ways to obtain SSR markers, namely: using published expressed sequence tags, using genome sequences that have been sequenced, and using transcriptome sequences. [0003] The relative species of Chinese cherry (Prunus pseudocerasus) include peach (P. persica), apricot (P. armeniaca), plum (P. salicina) and European sweet cherry (P. avium), among which the genome of peach has been seque...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 宗宇王月李永强朱友银郭卫东
Owner ZHEJIANG NORMAL UNIVERSITY
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